[Federal Register Volume 77, Number 172 (Wednesday, September 5, 2012)]
[Notices]
[Pages 54584-54597]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 2012-21849]
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DEPARTMENT OF HEALTH AND HUMAN SERVICES
National Institutes of Health
Final Action Under the NIH Guidelines for Research Involving
Recombinant DNA Molecules (NIH Guidelines)
SUMMARY: On March 4, 2009, the National Institutes of Health (NIH)
Office of Biotechnology Activities, Office of Science Policy (NIH/OBA)
published a proposal in the Federal Register (74 FR 9411) to revise the
NIH Guidelines in two regards. The first was to address biosafety
considerations for research with synthetic nucleic acids. The proposal
modified the scope of the NIH Guidelines specifically to cover certain
basic and clinical research with nucleic acid molecules created solely
by synthetic means. The second proposed revision was to modify the
criteria for determining whether an experiment to introduce drug
resistance into a microorganism must be reviewed by the Recombinant DNA
Advisory Committee (RAC) and approved by the NIH Director (as a Major
Action under Section III-A-1-a of the NIH Guidelines). Comments
submitted were discussed at the ``NIH Public Consultation on Proposed
Changes to the NIH Guidelines for Synthetic Nucleic Acids'' on June 23,
2009 (http://oba.od.nih.gov/rdna_rac/rac_pub_con.html''.
This notice sets forth final changes to the NIH Guidelines
regarding those two proposals. The scope of the NIH Guidelines is being
modified to cover certain classes of basic and clinical research with
synthetic nucleic acids while exempting others. As discussed herein,
the majority of research with synthetic nucleic acids that are not
designed to replicate does not raise significant biosafety concerns
that warrant oversight under the NIH Guidelines. Because of the
modification of the scope of the NIH Guidelines, the title of the NIH
Guidelines will be revised from NIH Guidelines for Research Involving
Recombinant DNA Molecules to NIH Guidelines for Research Involving
Recombinant or Synthetic Nucleic Acids Molecules.
These changes also clarify the criteria for determining whether an
experiment to introduce drug resistance into a microorganism raises
sufficient public health issues to warrant the experiment being
reviewed by the RAC and approved by the NIH Director under Section III-
A-1-a of the NIH Guidelines. While the current criteria for determining
whether an experiment requires review under Section III-A-1-a are being
retained, additional language is being added regarding the assessment
of whether a drug is therapeutically useful. In addition, NIH/OBA has
clarified that Institutional Biosafety Committees (IBCs) can consult
with NIH/OBA regarding a specific experiment that does not meet the
criteria for review under Section III-A-1-a but nonetheless raises
important public health issues. Finally, a section is added to give
NIH/OBA the authority
[[Page 54585]]
to approve new experiments utilizing the same drug resistance trait and
organism used in an experiment previously reviewed by the RAC and
approved by the NIH Director.
In March 2009, NIH/OBA also proposed changes to Section III-E-1 of
the NIH Guidelines, which sets containment for recombinant experiments
involving two-thirds or less of the genome of certain viruses in tissue
culture. In response to the comments on the proposed changes to Section
III-E-1, NIH/OBA revised the proposal and published a notice for
comment on April 22, 2010 (75 FR 21008). Comments received in response
to this notice were discussed at the June 16, 2010, public meeting of
the RAC and additional discussions of subsequent revisions to the
proposed changes took place at the June 7, 2011, meeting of the RAC. As
these changes are not yet finalized, NIH/OBA will move forward with the
other changes outlined below pending finalization of changes to Section
III-E-1.
DATES: These changes are effective March 5, 2013. All ongoing and
proposed experiments that will be newly subject to these amended NIH
Guidelines will need to be registered by the Principal Investigator
with the IBC by the effective date listed above. The six-month time
frame was deemed sufficient to allow institutions to develop new
procedures, as well as outreach and training for investigators whose
research will now be subject to the NIH Guidelines. While NIH/OBA does
not anticipate a significant increase in experiments subject to the NIH
Guidelines, it is important that institutions be afforded ample time to
implement effectively these changes.
FOR FURTHER INFORMATION CONTACT: If you have questions, or require
additional information about these proposed changes, please contact
NIH/OBA by e-mail at [email protected], by telephone at 301-496-9838, by
fax to 301-496-9839, or by mail to the Office of Biotechnology
Activities, National Institutes of Health, 6705 Rockledge Drive, Suite
750, MSC 7985, Bethesda, Maryland 20892.
SUPPLEMENTARY INFORMATION: As discussed in more detail in the March
2009 Federal Register notice, nucleic acid (NA) synthesis technology,
in combination with other rapidly evolving capabilities in the life
sciences, such as directed molecular evolution and viral reverse
genetics, has the potential to accelerate scientific discovery, yield
new therapeutics for disease, and facilitate the modification of
existing organisms or the creation of new organisms, including
pathogens.
The impetus for these changes to the NIH Guidelines is two-fold:
(1) Recognition that appropriate biosafety containment of an agent is
critical regardless of the technology used to generate that agent
(i.e., recombinant DNA or synthetic biology), and (2) a recommendation
from the National Science Advisory Board for Biosecurity (NSABB). The
NSABB was formed to advise the U.S. Government on strategies for
minimizing the potential for misuse of information, products, and
technologies from life sciences research, taking into consideration
both national security concerns and the needs of the research
community. In 2006, the NSABB published a report titled ``Addressing
Biosecurity Concerns Related to the Synthesis of Select Agents''
(available at http://oba.od.nih.gov/biosecurity/pdf/Final_NSABB_Report_on_Synthetic_Genomics.pdf).
In that report, the NSABB noted that practitioners of synthetic
genomics or researchers using synthetic nucleic acids in the emerging
field of synthetic biology are not necessarily biologists and,
therefore, may not have been trained in biosafety. These researchers
may be uncertain about how to conduct a risk assessment, as required
for research currently subject to the NIH Guidelines, and when to have
their work undergo review by an IBC. The NSABB report recommended that
the U.S. Government ``examine the language and implementation of
current biosafety guidance to ensure that such guidelines and
regulations provide adequate guidance for working with synthetically
derived DNA and are understood by all those working in areas addressed
by the guidelines.''
The recommendation on the need for examination of existing
biosafety guidance was accepted by the U.S. Government with the
understanding that implementation would be through examination and
modification of the NIH Guidelines, as appropriate. The changes to the
NIH Guidelines would then be cross-referenced in the joint publication
by the U.S. Centers for Disease Control and Prevention and NIH titled:
Biosafety in Microbiological and Biomedical Laboratories (BMBL)
(available at http://www.cdc.gov/biosafety/publications/bmbl5/index.htm).
As stated in the March 2009 Federal Register notice, these changes
were developed in consultation with the RAC. A total of 50 comments
were received in response to the March 2009 Federal Register notice
from individuals, academic and government researchers, private
pharmaceutical companies and trade organizations that represent the
biosafety community, researchers in gene and cell therapy, and
microbiologists. In addition, a day-long public discussion of the
proposed changes was held on June 23, 2009, in Arlington, Virginia. The
agenda and webcast of that meeting are available at the following URL:
http://oba.od.nih.gov/rdna_rac/rac_pub_con.html.
The NIH Guidelines currently apply to all recombinant DNA research
that is conducted at or sponsored by institutions that receive NIH
funding for any research involving recombinant DNA. In addition, some
federal agencies, including the U.S. Departments of Energy, Veterans
Affairs, and Agriculture, currently have policies in place stating that
all recombinant DNA research conducted by or funded by these agencies
must comply with the NIH Guidelines. While the NIH Guidelines may not
apply to all Government-funded and privately funded research, it may be
used as a tool for the entire research community to understand the
potential biosafety implications of this type of research.
Summary of Comments
All of the comments submitted in response to the Federal Register
notice are available for review on the NIH/OBA Web site at: http://oba.od.nih.gov/rdna_rac/rac_pub_con.html. The public comments
generally fell into two groups: (1) Comments on the proposed changes
regarding research with synthetic nucleic acids and (2) comments on the
proposed changes to Section III-A-1-a (experiments involving the
deliberate transfer of a drug resistance trait into microorganisms).
Overall, the comments favored modifying the scope of the NIH Guidelines
to include research with synthetic nucleic acids. As one commenter
noted, ``With the ability to chemically synthesize entire genes or
substantial portions of viral genomes, such synthetic entities would
have the potential to (1) Express proteins, (2) replicate in cells, and
(3) integrate into the host genome. As such, these entities warrant the
same scrutiny as traditional recombinant DNA with respect to studies
being conducted in [a] research laboratory and when being considered
for use in human subjects, and thus should be subject to NIH/OBA
registration and RAC review.'' However, there were concerns that the
proposed amendments would lead to oversight of the synthesis of small
nucleic acid primers used in basic research. This was a
misinterpretation of the proposed
[[Page 54586]]
changes; research with nucleic acids that are not in cells or organisms
is not subject to the NIH Guidelines and the proposed exemption for
non-replicating synthetic nucleic acids, discussed herein, would also
preclude these constructs from being subject to the NIH Guidelines.
Most of the comments regarding synthetic nucleic acids and the NIH
Guidelines focused on whether certain synthetic nucleic acids used in
human clinical trials should also be exempt from the NIH Guidelines and
in particular from the requirements for submission and review of human
gene transfer trials (as outlined in Appendix M of the NIH Guidelines).
These comments directly addressed a question posed in the March 2009
Federal Register: ``For human gene transfer research, are there classes
of non-replicating, synthetic molecules that should be exempt due to
lower potential risk (e.g. antisense RNA, RNAi)? If so, what criteria
should be applied to determine such classes?''
Many of the respondents to this question were involved in
developing such products to be used as therapeutics or represent
companies and investigators involved in such research. As discussed in
more detail herein, the respondents argued that small non-replicating
synthetic nucleic acids used as therapeutics are more akin to small
molecule drugs than traditional gene transfer agents. A session at the
June 23, 2009, public consultation focused on whether certain non-
replicating synthetic nucleic acids used in human clinical trials
should be exempted from the NIH Guidelines due to characteristics that
are distinct from recombinant molecules as currently defined in the NIH
Guidelines.
The second set of comments focused on the proposed changes to
Section III-A-1-a, which addresses certain experiments that involve the
introduction of drug resistance into microorganisms. The comments
uniformly disagreed with the proposed changes stating that the new
proposed criteria were too broad and would lead to federal review of
experiments that did not raise public health issues warranting
heightened scrutiny. Moreover, they stated that there is no evidence
that the current language had failed to serve the public health and
therefore the changes were not warranted given the potential problems
raised by expanding such review. As discussed herein, the III-A-1-a
language in the current NIH Guidelines (October 2011 version) will be
retained.
The following paragraphs review (1) The specific comments received
on each section of the NIH Guidelines, both the written comments and
those received at public meeting; (2) NIH/OBA's response to those
comments; and (3) the final changes to the NIH Guidelines.
Amendments to the NIH Guidelines
In order to ensure that biosafety considerations of synthetic
biology research are addressed appropriately, changes are being made to
the following sections of the NIH Guidelines:
Title of the NIH GuidelinesSection I. Scope of the NIH
Guidelines
Section I-B. Definition of Recombinant DNA
Section I-C. General Applicability
Section III-C. Experiments Involving the Deliberate Transfer of
Recombinant DNA,or DNA or RNA Derived from Recombinant DNA, into One
or More Human Research Participants
Section III-F. Exempt Experiments
Section IV-A. Policy
Section II-A-3. Comprehensive Risk Assessment
As discussed herein, the NIH Guidelines will no longer be limited
to oversight of research with recombinant nucleic acid molecules but
will also address research with certain synthetic nucleic acids.
Throughout the NIH Guidelines, the term ``recombinant DNA molecules''
will be replaced with ``recombinant or synthetic nucleic acids,'' which
will encompass research with either recombinant or synthetic or both
types of nucleic acids. This change will not be made to the name of the
Recombinant DNA Advisory Committee, although the Committee will provide
advice on both recombinant and synthetic nucleic acid research.
In addition to the changes being made specifically to address
research with synthetic nucleic acids, the following sections are also
being revised:
Section III-A-1. Major Actions under the NIH Guidelines
Section III-B. Experiments that Require NIH/OBA and
Institutional Biosafety Committee Approval before Initiation
Title of the NIH Guidelines
The title of the document will be changed from the NIH Guidelines
for Research Involving Recombinant DNA Molecules to the NIH Guidelines
for Research Involving Recombinant or Synthetic Nucleic Acid Molecules.
NIH received no comments regarding the proposed change to the title of
the NIH Guidelines.
Section I. Scope of the NIH Guidelines
To clarify the applicability of the NIH Guidelines to research
involving synthetic nucleic acids, modifications were proposed to
Section I, Scope of the NIH Guidelines. Section I-A (Purpose) of the
NIH Guidelines previously stated:
The purpose of the NIH Guidelines is to specify practices for
constructing and handling: (i) recombinant deoxyribonucleic acid
(DNA) molecules, and (ii) organisms and viruses containing
recombinant DNA molecules.
Section I-A was proposed to be changed to:
The purpose of the NIH Guidelines is to specify the practices
for constructing and handling: (i) recombinant nucleic acid
molecules, (ii) synthetic nucleic acid molecules, including those
wholly or partially containing functional equivalents of
nucleotides, and (iii) organisms and viruses containing such
molecules.
NIH/OBA received one comment regarding the use of the term
``constructing'' in reference to synthetic nucleic acids. The concern
was that the NIH Guidelines would govern the chemical synthesis of
nucleic acids. However, this language was not a revision to the
original scope of the NIH Guidelines. While the scope of the NIH
Guidelines has always referred to ``constructing'' or construction of
recombinant nucleic acids, the NIH Guidelines then exempts research
with nucleic acids that are not contained in cells, organisms, or
viruses. Therefore, the chemical synthesis of nucleic acids not placed
in cells, organisms, or viruses would likewise be exempt; the NIH
Guidelines will only apply once synthetic nucleic acids are placed in a
biological system.
NIH/OBA also received comments requesting a definition of the term
``functional equivalents of nucleotides.'' This term was intended to
capture synthetic nucleic acids that contain nucleotides that have been
chemically modified and do not have the same chemical structure as the
nucleotides in naturally occurring nucleic acids (see, for example, S.
Benner, Redesigning Genetics. Science. 306, 625-626 (2004)). For
clarity, the term ``functional equivalents'' has been changed to
``nucleotides that are chemically or otherwise modified but can base
pair with naturally occurring nucleic acid molecules.''
Thus, the amended Section 1-A Purpose will state:
Section 1-A. Purpose
The purpose of the NIH Guidelines is to specify the practices
for constructing and handling: (i) recombinant nucleic acid
molecules, (ii) synthetic nucleic acid molecules, including those
that are chemically or otherwise modified but can base pair with
naturally occurring nucleic acid molecules, and (iii) cells,
organisms, and viruses containing such molecules.
[[Page 54587]]
As a result of these modifications, the NIH Guidelines will apply
(unless otherwise exempted by other sections of the NIH Guidelines,
e.g. III-F) to both recombinant and synthetically derived nucleic
acids, including those that are chemically or otherwise modified
analogs of nucleotides (e.g. , morpholinos).
Section I-B. Definition of Recombinant Nucleic Acids
The current definition of a recombinant DNA molecule in the NIH
Guidelines (Section I-B) only explicitly refers to DNA and requires
that segments be joined, which may not need to occur in research with
synthetic nucleic acids. The revision to this section largely retains
the definition of recombinant DNA but also adds a definition for
synthetic nucleic acids that are created without joining segments of
nucleic acids.
Section I-B also contains a paragraph that states:
Synthetic DNA segments which are likely to yield a potentially
harmful polynucleotide or polypeptide (e.g. , a toxin or a
pharmacologically active agent) are considered as equivalent to
their natural DNA counterpart. If the DNA segment is not expressed
in vivo as a biologically active polynucleotide or polypeptide
product, it is exempt from the NIH Guidelines.
A second paragraph in the definition states:
Genomic DNA of plants and bacteria that have acquired a
transposable element, even if the latter was donated from a
recombinant vector no longer present, are not subject to the NIH
Guidelines unless the transposon itself contains recombinant DNA.
The final changes eliminate the first paragraph above, referring to
synthetic DNA segments, because the NIH Guidelines now specifically
includes an exemption for certain low-risk synthetic constructs (see
III-F-1). For consistency, the second paragraph on transposons was
moved to the portion of the NIH Guidelines that covers exemptions
(Section III-F). The NIH received no comments on eliminating the first
paragraph and moving the second paragraph; therefore these changes are
being implemented.
With respect to the definition of recombinant and synthetic nucleic
acids, NIH/OBA received several comments with suggestions to use a
single definition for recombinant and synthetic nucleic acids. NIH/OBA
considered these proposals carefully but decided instead to largely
retain the original definition of recombinant DNA, with clarification
that it applies to both DNA and RNA and to add a new definition of
synthetic nucleic acids. This was done because the definition of
recombinant DNA will not change with this revision to the NIH
Guidelines. As in the Scope section, the modification to the language
``functional equivalent'' will be included in the definition as well.
Section I-B is changed as follows:
Section I-B. Definition of Recombinant and Synthetic Nucleic
Acid Molecules:
In the context of the NIH Guidelines, recombinant and synthetic
nucleic acids are defined as:
(i) molecules that a) are constructed by joining nucleic acid
molecules and b) can replicate in a living cell, i.e. , recombinant
nucleic acids;
(ii) nucleic acid molecules that are chemically or by other
means synthesized or amplified, including those that are chemically
or otherwise modified but can base pair with naturally occurring
nucleic acid molecules, i.e. , synthetic nucleic acids; or
(iii) molecules that result from the replication of those
described in (i) or (ii) above.
Section I-C. General Applicability
In the March 2009 Federal Register notice, NIH/OBA stated that it
would change, throughout the NIH Guidelines, as appropriate, the term
``recombinant DNA molecules'' to ``recombinant and synthetic nucleic
acid molecules.'' NIH/OBA received a comment that this substitution
would imply that the NIH Guidelines only apply to research that uses
synthetic and recombinant nucleic acids together, not just recombinant
nucleic acid molecules or synthetic nucleic acid molecules alone. NIH/
OBA agrees with the comment on the original proposed language and
instead will replace, where appropriate recombinant DNA with
``recombinant or synthetic nucleic acid molecules'' to specify that the
section applies to research with recombinant or synthetic nucleic acids
or both. Section 1-C-1 currently states:
Section I-C. General Applicability
Section I-C-1. The NIH Guidelines are applicable to:
Section I-C-1-a. All recombinant DNA research within the United
States (U.S.) or its territories that is within the category of
research described in either Section I-C-1-a-(1) or Section I-C-1-a-
(2).
Section I-C-1-a-(1). Research that is conducted at or sponsored
by an institution that receives any support for recombinant DNA
research from NIH, including research performed directly by NIH. An
individual who receives support for research involving recombinant
DNA must be associated with or sponsored by an institution that
assumes the responsibilities assigned in the NIH Guidelines.
Section I-C-1-a-(2). Research that involves testing in humans of
materials containing recombinant DNA developed with NIH funds, if
the institution that developed those materials sponsors or
participates in those projects. Participation includes research
collaboration or contractual agreements, not mere provision of
research materials.
Section I-C-1-b. All recombinant DNA research performed abroad
that is within the category of research described in either Section
I-C-1-b-(1) or Section I-C-1-b-(2).
Section I-C-1-b-(1). Research supported by NIH funds.
Section I-C-1-b-(2). Research that involves testing in humans of
materials containing recombinant DNA developed with NIH funds, if
the institution that developed those materials sponsors or
participates in those projects. Participation includes research
collaboration or contractual agreements, not mere provision of
research materials.
Section I-C will now read:
Section I-C. General Applicability
Section I-C-1. The NIH Guidelines are applicable to:
Section I-C-1-a. All recombinant or synthetic nucleic acid
research within the United States (U.S.) or its territories that is
within the category of research described in either Section I-C-1-a-
(1) or Section I-C-1-a-(2).
Section I-C-1-a-(1). Research that is conducted at or sponsored
by an institution that receives any support for recombinant or
synthetic nucleic acid research from NIH, including research
performed directly by NIH. An individual who receives support for
research involving recombinant or synthetic nucleic acids must be
associated with or sponsored by an institution that assumes the
responsibilities assigned in the NIH Guidelines.
Section I-C-1-a-(2). Research that involves testing in humans of
materials containing recombinant or synthetic nucleic acids
developed with NIH funds, if the institution that developed those
materials sponsors or participates in those projects. Participation
includes research collaboration or contractual agreements, not mere
provision of research materials.
Section I-C-1-b. All recombinant or synthetic nucleic acid
research performed abroad that is within the category of research
described in either Section I-C-1-b-(1) or Section I-C-1-b-(2).
Section I-C-1-b-(1). Research supported by NIH funds.
Section I-C-1-b-(2). Research that involves testing in humans of
materials containing recombinant or synthetic nucleic acids
developed with NIH funds, if the institution that developed those
materials sponsors or participates in those projects. Participation
includes research collaboration or contractual agreements, not mere
provision of research materials.
[[Page 54588]]
Section III-C-1. Experiments Involving the Deliberate Transfer of
Recombinant DNA, or DNA or RNA Derived From Recombinant DNA, Into One
or More Human Research Participants
In March 2009, NIH/OBA proposed the following change to the
definition of human gene transfer:
For an experiment involving the deliberate transfer of
recombinant and/or synthetic nucleic acids into one or more human
research participants (human gene transfer), no research participant
shall be enrolled (see definition of enrollment in Section I-E-7)
until the RAC review process has been completed (see Appendix M-I-B,
RAC Review Requirements).
NIH/OBA had proposed exempting from the NIH Guidelines non-clinical
research with certain synthetic nucleic acids but did not propose to
extend that exemption to the use of these constructs in a clinical
setting. NIH/OBA noted that many gene transfer trials that are
currently subject to the NIH Guidelines use non-replicating recombinant
molecules because they are derived through recombinant technology which
involves replication. NIH/OBA proposed that there are shared safety
issues raised by clinical protocols that use synthetic non-replicating
nucleic acids and those that use non-replicating recombinant vectors.
The proposal to exempt basic research with non-replicating
synthetic nucleic acids but not to extend that exemption to human gene
transfer research was based on the differences in the potential health
risk from inadvertent exposure during basic or preclinical work versus
intentional exposure in a clinical setting. The doses and routes of
administration used in human gene transfer generally increase the
safety risks as compared to exposures that may occur in a basic
research setting. Moreover, the clinical safety risks to be considered
for human gene transfer are not limited to the replicative nature of
the vector but include transgene effects, risks of insertional
mutagenesis, immunological responses, and potential epigenetic changes.
Human gene transfer also raises scientific, medical, social, and
ethical considerations that warrant special attention and public
discussion.
NIH/OBA received a number of comments from industry, including
several comments from the Oligonucleotide Safety Working Group (OSWG),
which represents 70 pharmaceutical and regulatory professionals
involved in the clinical development of oligonucleotide-based
therapies. The OSWG stated that synthetic nucleic acid oligonucleotides
that are less than 100 nucleotides and are not delivered in a bacterial
or viral vector are more analogous to small molecule drugs than to the
agents currently used in human gene transfer. They noted that these
constructs can be distinguished from the recombinant agents currently
used in human gene transfer by their inability to integrate into the
genome or replicate in cells, their lack of a transgene that can be
transcribed into RNA or translated into a protein, and their transient
nature, i.e., they are degraded within days. They recognized that the
review of gene transfer protocols by the RAC is useful to address such
risks in gene transfer, but they did not believe that review should be
extended to these constructs merely because they are synthetic nucleic
acids. They noted that no significant safety issues have arisen in the
ongoing Phase I and Phase II clinical trials using short-interfering
RNA oligonucleotides (siRNAs). In addition to these trials, there is
significant interest in developing clinical applications directed at
microRNAs (miRNAs). For recent reviews of the field see K. Tiemann, J.
Rossi, RNAi-based therapeutics-current status, challenges and
prospects. EMBO Mol. Med. 1,142-151 (2009), and D. Grimm, M. A. Kay,
Therapeutic application of RNAi: is mRNA targeting finally ready for
prime time. The Journal of Clinical Investigation. 117(12), 3633-3641
(2007).
While this clinical data is reassuring, several preclinical
investigations raised important questions regarding the current
understanding about the mechanisms underlying the clinical action of
these constructs. For example, clinical trials using a siRNA against
vascular endothelial growth factor-A (VEGFA) or its receptor (VEGFR1)
in patients with blinding choroidal neovascularization (CNV) from age-
related macular degeneration have demonstrated promising results. The
hypothesis is that the siRNAs that are specific for VEGFA or its
receptor are responsible for the clinical responses seen. In 2008, M.E.
Kleinman, et al. found that a siRNA that did not specifically target
VEGFA or VEGFR1 could also suppress CNV in mice through an immune
response generated through toll-like receptors and induction of
interferon-gamma and interleukin-12 (see M.E., Kleinman, et al.,
Sequence- and target-independent angiogenesis suppression by siRNA via
TLR3. Nature. 452, 591-598 (2008)). In another study, investigators
developed anti-macrophage inhibitory factor (MIF) siRNAs designed to
block MIF expression in mammary adenocarcinoma cells (MCF-7). MIF is a
``pleiotropic cytokine with well described roles in cell proliferation,
tumorigenesis and angiogenesis'' (M.E. Armstrong, et al. , Small
Interfering RNAs Induce Macrophage Migration Inhibitory Factor
Production and Proliferation in Breast Cancer Cells via a Double
Stranded RNA-Dependent Protein Kinase-Dependent Mechanism. J. Imm.180,
7125-7133 (2008)). MIF has been shown to exert its actions through
activation of CD44 and enhanced CD44 activation has been shown to
promote breast cancer cell invasion. Unexpectedly, when these anti-MIF
siRNAs were delivered to MCF-7 cells, the result was increased MIF
production and an increase in proliferation of these cells.
In addition to questions regarding the mechanisms of action and
potential off target effects raised by these publications, the RAC
discussed whether administration of these synthetic RNAs could
potentially lead to long-term gene silencing and phenotypic changes. As
stated by the OSWG in their comments, one of the reasons for the RAC
oversight of recombinant research is to assess the potential for
alteration of a research participant's DNA, which could have unknown
and unintended consequences. Recent research indicates that siRNA and
miRNAs may be involved in long-term gene silencing (A. Verdel, et al.,
Common themes in siRNA-mediated epigenetic silencing pathways. Int. J.
Dev. Biol. 53, 245-257 (2009); D. H. Kim, et al. , MicroRNA-directed
transcriptional gene silencing in mammalian cells. PNAS. 105(42),
16230-16235 (2008)). The implications of these preliminary findings and
whether such effects on genes are fundamentally different than those
exerted by certain small molecules, for example histone deacetylation
inhibitors, remains an open question: It has been shown that histone
deacetylation can silence genes through chromatin modification and
deacetylation of the chromatin histone protein. Histone deacetylase
inhibitors are in development as potential cancer therapeutics (see
e.g. , A.A Lane, B.A. Chabner, Histone deacetylase inhibitors in cancer
therapy. J. Clin. Oncol. 27(32), 5459-68 (2009)).
After considering the comments by the OSWG and other interested
stakeholders, as well as the available literature, the RAC initially
recommended that NIH/OBA consider an exemption for certain well
characterized synthetic oligonucleotides, such as synthetic DNA
[[Page 54589]]
oligonucleotides that have been in clinical development for a number of
years and whose mechanism of action is well understood. The RAC had
reservations regarding extending that exemption to all synthetic RNA
oligonucleotides because of the emerging literature that raised
questions regarding our understanding of the potentially complex
biological pathways being targeted. Indeed certain pathways are highly
conserved across species and individual miRNAs have been shown to
suppress the production of hundreds of proteins (D. Baek, et al. The
impact of microRNAs on protein output. Nature. 455, 64-71(2008)).
Additionally, the RAC considered that review of clinical protocols that
administered RNA oligonucleotides without a vector would inform and
enhance the review of similar protocols that use vectors (e.g. , short
hairpin RNA (shRNA) expressed from a plasmid) and also inform the field
and promote the exchange of data that could enhance its development.
The RAC noted that this review might only be for several years until
more data were developed.
The RAC, however, continued to reflect upon the data and considered
additional stakeholder input. Further discussions were held with
leading experts on RNAi, including Noble Prize laureates Dr. Phillip
Sharp and Dr. Craig Mello. The RAC carefully considered the differences
between synthetic nucleic acids that are not delivered in vectors and
those delivered in bacterial or viral vectors, taking into account
their inability to replicate, integrate, or be transcribed or
translated. Finally, given the uncertain significance of preclinical
data in the absence of adverse effects in the ongoing clinical trials,
the RAC concluded that oversight is not warranted at this time. NIH/OBA
concurs with this assessment, and the NIH Guidelines will only apply to
recombinant constructs that are currently covered by the NIH Guidelines
and those synthetic constructs that are equivalent to their recombinant
counterparts, i.e. synthetic investigational agents that share the same
characteristics as recombinant gene transfer constructs. However, in
light of some unresolved outstanding questions regarding the mechanisms
of actions of synthetic nucleic acids used clinically, including the
potential for epigenetic changes, the RAC recommended NIH/OBA convene a
meeting to further explore these questions. NIH/OBA hosted this meeting
on December 15-16, 2011. (The agenda and slide presentations are
available at: http://oba.od.nih.gov/rdna/rdna_symposia.html.)
Therefore, Section III-C-1 will be revised as follows:
Section III-C-1. Experiments Involving the Deliberate Transfer
of Recombinant or Synthetic Nucleic Acid Molecules, or DNA or RNA
Derived from Recombinant or Synthetic Nucleic Acid Molecules, into
One or More Human Research Participants
Human gene transfer is the deliberate transfer into human
research participants of either:
1. Recombinant nucleic acid molecules, or DNA or RNA derived
from recombinant nucleic acid molecules, or
2. Synthetic nucleic acid molecules, or DNA or RNA derived from
synthetic nucleic acid molecules, that meet any one of the following
criteria:
a. Contain more than 100 nucleotides; or
b. Possess biological properties that enable integration into
the genome (e.g., cis elements involved in integration); or
c. Have the potential to replicate in a cell; or
d. Can be translated or transcribed.
No research participant shall be enrolled (see definition of
enrollment in Section 1-E-7) until the RAC review process has been
completed (see Appendix M-1-B, RAC Review Requirements).
Section III-F. Exempt Experiments
Modifications were proposed to augment or clarify experiments that
are exempt from the NIH Guidelines (III-F). Certain nucleic acid
molecules are exempt from the NIH Guidelines under Section III-F
because (1) their introduction into a biological system is not expected
to present a biosafety risk that requires review by an IBC, or (2) the
introduction of these nucleic acid molecules into biological systems
would be akin to processes of nucleic acid transfer that already occur
in nature, so that the appropriate biosafety practices would be the
same as those used for the natural organism and/or would be covered by
other guidances.
As stated in the March 2009 Federal Register notice, with the
exception of the new proposed Section III-F-1 discussed below, the
exemptions from the current NIH Guidelines (October 2011) have been
preserved with minor modifications. The addition of research with
synthetic nucleic acids to the NIH Guidelines does not warrant
modification of most of these exemptions except to extend them to
synthetic constructs.
To emphasize that research exempt from the NIH Guidelines may still
have biosafety considerations and that other standards of biosafety may
apply, a modification is being made to the introductory language for
this section. Section III-F currently states:
The following recombinant DNA molecules are exempt from the NIH
Guidelines and registration with the Institutional Biosafety
Committee is not required.
This portion is amended to read:
The following recombinant or synthetic nucleic acid molecules
are exempt from the NIH Guidelines and registration with the
Institutional Biosafety Committee is not required; however, other
federal and state standards of biosafety may still apply to such
research (for example, the Centers for Disease Control and
Prevention (CDC)/NIH publication Biosafety in Microbiological and
Biomedical Laboratories).
Section III-F-1. Exempt Experiments
A new entry under Section III-F was proposed to exempt from the NIH
Guidelines synthetic nucleic acids that cannot replicate unless they
are administered to one or more human research participant(s) (see
Section III-C-1). This exemption was proposed so that the NIH
Guidelines apply to synthetic nucleic acid research in a manner
consistent with the current oversight of basic and preclinical
recombinant DNA research. Currently oversight is limited to recombinant
molecules that replicate or are derived from such molecules. The added
section exempts basic, non-clinical research with synthetic nucleic
acids that cannot replicate or are not derived from molecules that can
replicate. The biosafety risks of using such constructs in basic and
preclinical research are likely low. If a nucleic acid is incapable of
replicating in a cell, any toxicity associated with that nucleic acid
should be confined to that particular cell or organism, and spread to
neighboring cells or organisms should not occur to any appreciable
degree. This type of risk is analogous to that observed with chemical
exposures, although nucleic acids are generally far less toxic than
most chemicals.
NIH/OBA received a number of comments on this proposed exemption.
Most of the comments questioned whether this exemption should be
extended to certain non-replicating nucleic acids used in human gene
transfer because such constructs are likely to pose quantitatively
different risks than vector-based gene transfer. The response to these
comments is articulated in the prior section of this notice that
focuses on Section III-C-1.
With respect to basic research, NIH/OBA received comments
questioning whether all non-replicating synthetic nucleic acids used in
basic research pose sufficiently low biosafety risks to be exempt from
the NIH Guidelines. Concerns were also raised about the use of
synthetic non-replicating, integrating
[[Page 54590]]
viral vectors, such as lentiviral vectors, which could result in
persistent transgene expression and have the potential to induce
insertional oncogenesis. Non-replicating synthetic cassettes for toxins
were also identified as raising potential biosafety risks as were
oncogenes. In addition, clarification was sought regarding what was
meant by the term ``replication.'' For example, would the following be
considered replicating nucleic acids: (1) Plasmids lacking sequences to
replicate in eukaryotic cells or (2) complementary DNAs (cDNAs) of
positive strand RNA viruses, in which cDNA is not replicated but is
transcribed into viral RNAs? In addition, another commenter asked why
the exemption was limited to synthetic nucleic acids rather than all
nucleic acids.
NIH/OBA carefully considered all of these comments. With respect to
making this exemption apply generally to all nucleic acid constructs,
recombinant and synthetic, NIH/OBA notes that the definition of
recombinant DNA molecules, which remains unchanged, only includes
molecules that can replicate in a living cell or molecules that result
from the replication of those described above. Therefore, to include
them in the exemption under III-F-1 would be redundant, as this
exemption only applies to nucleic acids that cannot replicate and are
not derived from those that can replicate. NIH/OBA acknowledges that
research with an integrating vector could raise biosafety
considerations even if the vector does not replicate. With respect to
toxins, a non-replicating expression cassette can only express the
toxin in a single cell and the toxin cannot spread from cell to cell,
thereby limiting its toxic effect. Nonetheless, NIH/OBA agrees that
constructs expressing toxins that are currently reviewed under Section
III-B-1, Experiments Involving the Cloning of Toxin Molecules with LD50
of Less Than 100 Nanograms per Kilogram Body Weight, should remain
subject to the NIH Guidelines. Indeed, under the current NIH
Guidelines, even if an experiment falls under a Section III-F
exemption, it may still be subject to review under Section III-B-1. For
clarity, NIH/OBA therefore decided to specify that toxin-producing
expression cassettes that would fall under Section III-B-1 will not be
exempt under III-F.
Synthetic constructs that have the potential to integrate will not
likewise be exempted because they could inadvertently activate an
oncogene, or an integrating sequence containing an oncogene could
inadvertently be integrated into a cell and persist and transform that
cell and its progeny.
In the March 2009 Federal Register notice, Section III-F-1 was
written so as to exempt from the NIH Guidelines ``Synthetic nucleic
acids that cannot replicate, and that are not deliberately transferred
into one or more human research participants (Section III-C and
Appendix M).'' To clarify the interpretation of ``replicating,'' the
language has been changed to match more closely that of the definition
of recombinant DNA, ``cannot replicate in a living cell.'' This change
is to make it clear that it is the ability to replicate in any cell
type that determines whether the research is subject to the NIH
Guidelines (i.e. , plasmids that can replicate in bacteria would be
subject to the NIH Guidelines even if in eukaryotic cells). To address
the cDNA of positive strand RNA viruses, the language has been changed
to ``cannot replicate or generate nucleic acids that can replicate in a
living cell.'' In addition, to make it clear that a synthetic
replication incompetent virus is not exempt under this section of the
NIH Guidelines, a parenthetical has been added to clarify that this
section is meant to exempt only research with small synthetic
oligonucleotides and expression cassettes, not synthetic viruses or
bacteria that cannot replicate because of omission of one or more
genes.
Section III-F-1 is changed to exempt the following experiments:
Section III-F-1. Those synthetic nucleic acids that: (1) Can
neither replicate nor generate nucleic acids that can replicate in
any living cell (e.g. , oligonucleotides or other synthetic nucleic
acids that do not contain an origin of replication or contain
elements known to interact with either DNA or RNA polymerase), and
(2) are not designed to integrate into DNA, and (3) do not produce a
toxin that is lethal for vertebrates at an LD50 of less than 100
nanograms per kilogram body weight. If a synthetic nucleic acid is
deliberately transferred into one or more human research
participants and meets the criteria of Section III-C, it is not
exempt under this Section.
Section III-F-2. Exempt Experiments
Section III-F-1 will now be renumbered to III-F-2 and is amended to
clarify that replicating nucleic acids that are not in cells,
organisms, or viruses are exempt. The current NIH Guidelines only
mentions organisms and viruses, and for clarity the term ``cells'' has
been added. In addition, if a molecule is modified to facilitate entry
into a cell, this will also not be exempt. Nucleic acids that are not
in a biological system that will permit replication and that have not
been modified to enable improved penetration of cell membranes are
unlikely to have associated biosafety risks. NIH/OBA received no
comments on this change.
The current Section III-F-1 states: ``Those that are not in
organisms or viruses.''
Section III-F-1 is re-numbered to III-F-2 and will exempt the
following experiments:
Section III-F-2. Those that are not in organisms, cells, or
viruses and that have not been modified or manipulated (e.g.,
encapsulated into synthetic or natural vehicles) to render them
capable of penetrating cellular membranes.
Sections III-F-3 through III-F-7
Revised Sections III-F-3 through III-F-7 retain exemptions that
were in the current version of NIH Guidelines (October 2011) with minor
revisions. There were no comments to the minor changes made in Sections
III-F-3 through III-F-7. The following changes will be made for these
Section III-F exemptions.
Section III-F-3. Exempt Experiments
Section III-F-2 exempts nucleic acid sequences that are essentially
copies of those found in nature. The language has been modified as
discussed in the March 2009 Federal Register notice by limiting this
exemption to those nucleic acid sequences that exist contemporaneously
in nature. Research in the lab with nucleic acid sequences for
organisms that do not currently exist in nature, for example, an
identical copy of the 1918 H1N1 influenza virus would not be exempt.
Section III-F-2 will be re-numbered to III-F-3 and will exempt the
following experiments:
Section III-F-3. Those that consist solely of the exact
recombinant or synthetic nucleic acid sequence from a single source
that exists contemporaneously in nature.
Section III-F-4. Exempt Experiments
The current Section III-F-3 exempts nucleic acids that are being
propagated only in a prokaryotic host that is either the natural host
or a closely related strain of the natural host. Again such constructs
may already exist outside of a laboratory. It is renumbered to Section
III-F-4 and no amendment to the language is made. It exempts the
following experiments:
Section III-F-4. Those that consist entirely of nucleic acids
from a prokaryotic host, including its indigenous plasmids or
viruses when propagated only in that host (or a closely related
strain of the same species), or when transferred to another host by
well established physiological means.
Section III-F-5: Exempt Experiments
The current Section III-F-4 exempts nucleic acids that are being
propagated
[[Page 54591]]
in a eukaryotic host that is either the natural host or closely related
strain of the natural host. Section III-F-4 is renumbered to Section
III-F-5 and no amendment to the language is made. The following
experiments are exempt per this section.
Section III-F-5. Those that consist entirely of nucleic acids
from a eukaryotic host including its chloroplasts, mitochondria, or
plasmids (but excluding viruses) when propagated only in that host
(or a closely related strain of the same species).
Section III-F-6. Exempt Experiments
Research that falls under Section III-F-6 (formerly Section III-F-
5) is exempt because the manipulation of these nucleic acids in a
laboratory setting would be equivalent to processes that occur in
nature when certain organisms exchange genetic material via
physiological processes (e.g. , bacterial conjugation). It is limited
to those organisms, as specified in Appendices A-I through A-VI, that
are already known to exchange DNA in nature. The current Section III-F-
5 is renumbered to Section III-F-6 and no amendment to the language is
made. The following experiments are exempt per this section.
Section III-F-6. Those that consist entirely of DNA segments
from different species that exchange DNA by known physiological
processes, though one or more of the segments may be a synthetic
equivalent. A list of such exchangers will be prepared and
periodically revised by the NIH Director with advice of the RAC
after appropriate notice and opportunity for public comment (see
Section IV-C-1-b-(1)-(c), Major Actions). See Appendices A-I through
A-VI, Exemptions under Section III-F-6--Sublists of Natural
Exchangers, for a list of natural exchangers that are exempt from
the NIH Guidelines.
Additionally, Appendix A will be amended to reference Section III-
F-6 rather than III-F-5.
Section III-F-7. Exempt Experiments
Research that falls under the proposed Section III-F-7 exemption
also involves a natural physiological process, i.e. transposition.
Transposons are nucleic acid molecules that exist in a wide variety of
organisms from bacteria to humans. These molecules have the ability to
move from one portion of an organism's genome to another. This new
Section of III-F captures what was previously an exemption to the
definition of a recombinant DNA molecule in the NIH Guidelines (Section
I-B). Unless a transposon has been modified to be a recombinant
molecule, genomic DNA that has acquired a transposon is not subject to
the NIH Guidelines. Transposons that have not been modified by the
insertion of recombinant or synthetic DNA are equivalent to what exists
in nature and the process occurs naturally outside of a laboratory
setting. The language from the definition of recombinant DNA (Section
I-B) is being moved to this Section so that the definition of
recombinant and synthetic nucleic acids found in Section I-B is solely
a definition and does not include exemptions. The exemption described
in Section I-B previously stated, ``Genomic DNA molecules of plants and
bacteria that have acquired a transposable element, even if the latter
was donated from a recombinant vector no longer present, are not
subject to the NIH Guidelines unless the transposon itself contains
recombinant DNA.'' The exemption language has been simplified to make
it clear that unmodified transposons used in research are not subject
to the NIH Guidelines even if derived from a recombinant or synthetic
system. In addition, the reference to only plants and bacteria has been
removed since it is now known that transposons are also found in
animals. Section III-F-7 will exempt the following experiments:
Section III-F-7. Those genomic DNA molecules that have acquired
a transposable element, provided the transposable element does not
contain any recombinant and/or synthetic DNA.
Section III-F-8. Exempt Experiments
The current Section III-F-6 provides a mechanism by which other
experiments that do not raise significant biosafety risks can be
exempted from the NIH Guidelines after review by the RAC and approval
by the NIH Director. The language has not been amended but, due to the
insertion of two additional exemptions, it is being renumbered to
Section III-F-8 and will exempt the following experiments:
Section III-F-8. Those that do not present a significant risk to
health or the environment (see Section IV-C-1-b-(1)-(c), Major
Actions), as determined by the NIH Director, with the advice of the
RAC, and following appropriate notice and opportunity for public
comment. See Appendix C, Exemptions under Section III-F-8 for other
classes of experiments which are exempt from the NIH Guidelines.
Additionally, Appendix C will be amended to reference Section III-
F-8 rather than III-F-6.
Section IV-A. Policy
Section IV-A addresses the roles and responsibilities of local
institutions and investigators in implementing the NIH Guidelines. It
contains a general policy statement that acknowledges the inability of
the NIH Guidelines to address specifically all conceivable research or
emerging techniques and therefore states that researchers and
institutions should adhere to ``the intent of the NIH Guidelines as
well as to their specifics.'' NIH/OBA received no comments on the
proposed changes, which emphasize that the NIH Guidelines are expected
to be modified to address new developments in research or scientific
techniques. In addition, in rewriting this section of the NIH
Guidelines, NIH/OBA has removed the sentence ``[G]eneral recognition of
institutional authority and responsibility properly establishes
accountability for safe conduct of the research at the local level,''
since the previous sentences adequately explains that the institution
is accountable for implementation of the NIH Guidelines. Section IV-A
currently states:
The safe conduct of experiments involving recombinant DNA
depends on the individual conducting such activities. The NIH
Guidelines cannot anticipate every possible situation. Motivation
and good judgment are the key essentials to protection of health and
the environment. The NIH Guidelines are intended to assist the
institution, Institutional Biosafety Committee, Biological Safety
Officer, and the Principal Investigator in determining safeguards
that should be implemented. The NIH Guidelines will never be
complete or final since all conceivable experiments involving
recombinant DNA cannot be foreseen. Therefore, it is the
responsibility of the institution and those associated with it to
adhere to the intent of the NIH Guidelines as well as to their
specifics. Each institution (and the Institutional Biosafety
Committee acting on its behalf) is responsible for ensuring that all
recombinant DNA research conducted at or sponsored by that
institution is conducted in compliance with the NIH Guidelines.
General recognition of institutional authority and responsibility
properly establishes accountability for safe conduct of the research
at the local level. The following roles and responsibilities
constitute an administrative framework in which safety is an
essential and integral part of research involving recombinant DNA
molecules. Further clarifications and interpretations of roles and
responsibilities will be issued by NIH as necessary.
Section IV-A is amended to read:
The safe conduct of experiments involving recombinant or
synthetic nucleic acid molecules depends on the individual
conducting such activities. The NIH Guidelines cannot anticipate
every possible situation. Motivation and good judgment are the key
essentials to protection of health and the environment. The NIH
Guidelines are intended to assist the institution, Institutional
Biosafety Committee, Biological Safety Officer, and the Principal
Investigator in determining safeguards that should be implemented.
The NIH Guidelines will never be complete or final since all
experiments
[[Page 54592]]
involving recombinant or synthetic nucleic acid molecules cannot be
foreseen. The utilization of new genetic manipulation techniques may
enable work previously conducted using recombinant means to be
accomplished faster, more efficiently, or at larger scale. These
techniques have not yet yielded organisms that present safety
concerns that fall outside the current risk assessment framework
used for recombinant nucleic acid research. Nonetheless, an
appropriate risk assessment of experiments involving these
techniques must be conducted taking into account the way these
approaches may alter the risk assessment. As new techniques develop,
the NIH Guidelines should be periodically reviewed to determine
whether and how such research should be explicitly addressed.
It is the responsibility of the institution and those associated
with it to adhere to the intent of the NIH Guidelines as well as to
its specifics. Therefore, each institution (and the Institutional
Biosafety Committee acting on its behalf) is responsible for
ensuring that all research with recombinant or synthetic nucleic
acid molecules conducted at or sponsored by that institution is
conducted in compliance with the NIH Guidelines. The following roles
and responsibilities constitute an administrative framework in which
safety is an essential and integral part of research involving
recombinant or synthetic nucleic acid molecules. Further
clarifications and interpretations of roles and responsibilities
will be issued by NIH as necessary.
Section II-A-3. Comprehensive Risk Assessment
Currently, the risk assessment framework of the NIH Guidelines uses
the Risk Group (RG) of the parent organism as a starting point for
determining the necessary containment level. For example, genetic
modifications of a Risk Group 3 organism (defined as agents that are
associated with serious or lethal human disease for which preventive or
therapeutic interventions may be available) would generally be carried
out at Biosafety Level 3 (BL3) containment, but the containment level
might be raised or lowered depending on the specific construct and the
experimental manipulations. The RAC concluded that the current risk
assessment framework under the NIH Guidelines can be effectively
applied to assess the biosafety risks of experiments with synthetic
nucleic acids. However, additional language was proposed to provide
further guidance for evaluating synthetic biology research, which has
the potential to create complex, novel organisms for which
identification of a parent organism may be more difficult or may not be
as relevant to the risk assessment as it is with more traditional
recombinant organisms. The risk assessment may also be complicated by
the limitations in predicting function from sequence(s), as recently
addressed in a report by the Committee on Scientific Milestones for the
Development of Gene-Sequence-Based Classification System for the
Oversight of Select Agents, National Research Council, Sequence-Based
Classification of Select Agents: A Brighter Line, ISBN-10: 0-309-15904-
0. Further complications may also result from synergistic effects
caused by combining sequences from different sources in a novel
context.
NIH/OBA received one comment on its proposed revisions to Section
II-A-3. The comment asked for clarification of the meaning of the term
``chimera'' because it is not currently used in the NIH Guidelines. The
term was meant to capture the concept that with the advent of more
sophisticated synthetic techniques, a complex organism may be created
using nucleic acid sequences from multiple sources. For clarity, this
wording will be used in lieu of the term ``chimera.''
Section II-A-3 Comprehensive Risk Assessment currently states:
In deciding on the appropriate containment for an experiment,
the initial risk assessment from Appendix B, Classification of Human
Etiologic Agents on the Basis of Hazard, should be followed by a
thorough consideration of the agent itself and how it is to be
manipulated. Factors to be considered in determining the level of
containment include agent factors such as: Virulence, pathogenicity,
infectious dose, environmental stability, route of spread,
communicability, operations, quantity, availability of vaccine or
treatment, and gene product effects such as toxicity, physiological
activity, and allergenicity. Any strain that is known to be more
hazardous than the parent (wild-type) strain should be considered
for handling at a higher containment level. Certain attenuated
strains or strains that have been demonstrated to have irreversibly
lost known virulence factors may qualify for a reduction of the
containment level compared to the Risk Group assigned to the parent
strain (see Section V-B, Footnotes and References of Sections I-IV).
A final assessment of risk based on these considerations is then
used to set the appropriate containment conditions for the
experiment (see Section II-B, Containment). The containment level
required may be equivalent to the Risk Group classification of the
agent or it may be raised or lowered as a result of the above
considerations. The Institutional Biosafety Committee must approve
the risk assessment and the biosafety containment level for
recombinant DNA experiments described in Sections III-A, Experiments
that Require Institutional Biosafety Committee Approval, RAC Review,
and NIH Director Approval Before Initiation; III-B, Experiments that
Require NIH/OBA and Institutional Biosafety Committee Approval
Before Initiation; III-C, Experiments that Require Institutional
Biosafety Committee and Institutional Review Board Approvals and
NIH/OBA Registration Before Initiation; III-D, Experiments that
Require Institutional Biosafety Committee Approval Before
Initiation.
Careful consideration should be given to the types of
manipulation planned for some higher Risk Group agents. For example,
the RG2 dengue viruses may be cultured under the Biosafety Level 2
(BL2) containment (see Section II-B); however, when such agents are
used for animal inoculation or transmission studies, a higher
containment level is recommended. Similarly, RG3 agents such as
Venezuelan equine encephalomyelitis and yellow fever viruses should
be handled at a higher containment level for animal inoculation and
transmission experiments.
Individuals working with human immunodeficiency virus (HIV),
hepatitis B virus (HBV) or other bloodborne pathogens should consult
the applicable Occupational Safety and Health Administration (OSHA)
regulation, 29 CFR 1910.1030, and OSHA publications, e.g., OSHA
3186-06R (2003 revised). BL2 containment is recommended for
activities involving all blood-contaminated clinical specimens, body
fluids, and tissues from all humans, or from HIV-or HBV-infected or
inoculated laboratory animals. Activities such as the production of
research-laboratory scale quantities of HIV or other bloodborne
pathogens, manipulating concentrated virus preparations, or
conducting procedures that may produce droplets or aerosols, are
performed in a BL2 facility using the additional practices and
containment equipment recommended for BL3. Activities involving
industrial scale volumes or preparations of concentrated HIV are
conducted in a BL3 facility, or BL3 Large Scale if appropriate,
using BL3 practices and containment equipment.
Exotic plant pathogens and animal pathogens of domestic
livestock and poultry are restricted and may require special
laboratory design, operation and containment features not addressed
in Biosafety in Microbiological and Biomedical Laboratories (see
Section V-C, Footnotes and References of Sections I through IV). For
information regarding the importation, possession, or use of these
agents see Section V-G and V-H, Footnotes and References of Sections
I through IV.
The first paragraph is being revised to clarify that the assignment
of an organism to a Risk Group in Appendix B, Classification of Human
Etiologic Agents on the Basis of Hazard, is based on a risk assessment
and identification of the Risk Group of the parent organism. The first
paragraph is amended as follows:
In deciding on the appropriate containment for an experiment,
the first step is to assess the risk of the agent itself. Appendix
B, Classification of Human Etiologic Agents on the Basis of Hazard,
classifies agents into Risk Groups based on an assessment of their
ability to cause disease in humans and the available treatments for
such disease. Once the Risk Group of the agent is identified, this
should be followed
[[Page 54593]]
by a thorough consideration of how the agent is to be manipulated.
Factors to be considered in determining the level of containment
include agent factors such as: Virulence, pathogenicity, infectious
dose, environmental stability, route of spread, communicability,
operations, quantity, availability of vaccine or treatment, and gene
product effects such as toxicity, physiological activity, and
allergenicity. Any strain that is known to be more hazardous than
the parent (wild-type) strain should be considered for handling at a
higher containment level. Certain attenuated strains or strains that
have been demonstrated to have irreversibly lost known virulence
factors may qualify for a reduction of the containment level
compared to the Risk Group assigned to the parent strain (see
Section V-B, Footnotes and References of Sections I-IV).
The following new paragraphs will then be inserted:
While the starting point for the risk assessment is based on the
identification of the Risk Group of the parent agent, as technology
moves forward, it may be possible to develop an organism containing
genetic sequences from multiple sources such that the parent agent
may not be obvious. In such cases, the risk assessment should
include at least two levels of analysis. The first involves a
consideration of the Risk Groups of the source(s) of the sequences
and the second involves an assessment of the functions that may be
encoded by these sequences (e.g., virulence or transmissibility). It
may be prudent to first consider the highest Risk Group
classification of all agents that are the source of sequences
included in the construct. Other factors to be considered include
the percentage of the genome contributed by each parent agent and
the predicted function or intended purpose of each contributing
sequence. The initial assumption should be that all sequences will
function as they did in the original host context.
The Principal Investigator and Institutional Biosafety Committee
must also be cognizant that the combination of certain sequences in
a new biological context may result in an organism whose risk
profile could be higher than that of the contributing organisms or
sequences. The synergistic function of these sequences may be one of
the key attributes to consider in deciding whether a higher
containment level is warranted, at least until further assessments
can be carried out. A new biosafety risk may occur with an organism
formed through combination of sequences from a number of organisms
or due to the synergistic effect of combining transgenes that
results in a new phenotype.
A final assessment of risk based on these considerations is then
used to set the appropriate containment conditions for the
experiment (see Section II-B, Containment). The appropriate
containment level may be equivalent to the Risk Group classification
of the agent or it may be raised or lowered as a result of the above
considerations. The Institutional Biosafety Committee must approve
the risk assessment and the biosafety containment level for
recombinant or synthetic nucleic acid experiments described in
Sections III-A, Experiments that Require Institutional Biosafety
Committee Approval, RAC Review, and NIH Director Approval Before
Initiation; III-B, Experiments that Require NIH/OBA and
Institutional Biosafety Committee Approval Before Initiation; III-C,
Experiments that Require Institutional Biosafety Committee and
Institutional Review Board Approvals and NIH/OBA Registration Before
Initiation; and III-D, Experiments that Require Institutional
Biosafety Committee Approval Before Initiation.
Section III-A-1. Major Actions under the NIH Guidelines
In reviewing the NIH Guidelines and the different levels of review
required for each category of experiment, the RAC determined that it is
important also to evaluate the class of experiments that require the
highest level of review: Both RAC review and NIH Director approval. In
doing so, it was determined that the language for Section III-A-1-a of
the NIH Guidelines (research involving the introduction of drug
resistance into a microorganism) may not capture all of the experiments
that warrant this heightened review. Moreover, given the change in the
use of antibiotics and the public health problems raised by the
emergence of multidrug resistant bacterial strains, clearly defining
those experiments that require heightened review is a public health
priority.
Section III-A-1-a currently states:
The deliberate transfer of a drug resistance trait to
microorganisms that are not known to acquire the trait naturally
(see Section V-B, Footnotes and References of Sections I-IV), if
such acquisition could compromise the use of the drug to control
disease agents in humans, veterinary medicine, or agriculture, will
be reviewed by RAC.
In March 2009, NIH/OBA proposed to remove the phrase not known to
acquire the trait naturally in order to allow some flexibility in
review of experiments that may raise public health concern but for
which there may be low levels of antibiotic resistance in the
community. For example, only a small number of vancomycin-resistant
Staphylococcus aureus strains have been isolated (B.P. Howden, et al. ,
Reduced Vancomycin Susceptibility in Staphylococcus aureus, including
Vancomycin-Intermediate and Heterogeneous Vancomycin-Intermediate
Strains: Resistance Mechanisms, Laboratory Detection and Clinical
Implications. Clinical Microbiology Reviews. 32(1), 99-139 (2010)).
However, as there are only a limited number of antibiotics with which
to treat these multidrug resistant S. aureus strains, the use of
vancomycin resistance as a marker could raise public health concerns.
Another example would be the use of ciprofloxacin resistance as a
marker for Neisseria meningitidis. Again, there are a small number of
documented cases of resistance, but ciprofloxacin remains the primary
drug for post-exposure prophylaxis (H.M. Wu, et al., Emergence of
Ciprofloxacin-resistant Neisseria meningitides in North America. N.
Engl. J. Med. 360(9), 886-92 (2009)).
In the March 2009 Federal Register notice, Section III-A-1-a was
proposed to be amended as follows:
The deliberate transfer of a drug resistance trait to
microorganisms, if such acquisition could compromise the ability to
treat or manage disease agents in human and veterinary medicine, or
agriculture will be reviewed by RAC. Even if an alternative drug or
drugs exist for the control or management of disease, it is
important to consider how the research might affect the ability to
control infection in certain groups or subgroups by putting them at
risk of developing an infection by such microorganism for which
alternative treatments may not be available. Affected groups or
subgroups may include, but are not limited to: children, pregnant
women, and people who are allergic to effective alternative
treatments, immunocompromised or living in countries where the
alternative effective treatment is not readily available.
In response to this proposed change in the language to Section III-
A-1, NIH/OBA received a total of 36 written comments. Most either
specifically noted their concurrence with comments from the American
Society for Microbiology (ASM) or substantively concurred with ASM's
comment. ASM commented that based on their interpretation of the
proposed language the net effect would be to broaden substantially the
scope of research that would be subject to the requirements of Section
III-A-1-a and ``have a chilling impact on microbiological research
where antibiotic resistance is routinely used in molecular and genetic
studies.'' The ASM did agree that whether an organism is ``known to
acquire the trait naturally'' is not always the critical factor in
evaluating the safety of the experiment. ASM further stated that
broadening the range of concern to include consideration of possible
rare uses of an antibiotic that is not the ``drug of choice'' will only
confound the work of the IBCs.
Other commenters noted that it was the overuse and likely misuse
use of antibiotics throughout the world that pose a much greater and
better documented public health threat through the development of
highly resistant organisms that are capable of surviving outside of a
laboratory. They noted that this threat is distinct from the laboratory
setting as many laboratory-generated strains may not have a
[[Page 54594]]
selective advantage outside the laboratory and, even if there were
inadvertent release, may not become a public health risk. Some comments
suggested adding qualifiers to narrow the scope of the proposed
section. For example, one commenter suggested the addition of the word
``reasonably'' to the concept of whether the transfer of drug
resistance could compromise the ability to treat disease. Another
commenter suggested that a list of criteria be developed that could be
considered when a determination is made as to whether the transfer of a
drug resistance trait could compromise public health. An additional
commenter suggested that a list of ``acceptable'' transfers of drug
resistance be incorporated into the NIH Guidelines.
Other comments revealed some potential misinterpretation of what
constitutes research that falls under Section III-A-1-a. For
clarification, NIH/OBA notes that transfer of a drug resistance trait
to any non-pathogenic organism is not subject to the requirements of
Section III-A-1-a of the NIH Guidelines, and transfer of resistance to
a drug that is not currently used to treat disease caused by a
pathogenic organism is not subject to review under Section III-A-1-a.
These experiments, however, may be subject to other portions of the NIH
Guidelines.
The changes proposed in the March 2009 Federal Register notice were
further discussed at the public consultation on June 23, 2009. The
panel of experts generally agreed that public health concerns may be
raised by the use of certain antibiotic markers in pathogens that have
resistance to a number of antibiotics, for example the use of
vancomycin resistance as a marker in S. aureus. However, they concluded
that these concerns could be adequately addressed by the IBC by
requiring appropriate containment. The experts at the June 23, 2009,
meeting agreed with ASM's observation that the safety of an experiment
is not dictated solely by whether the organism can naturally acquire
the resistance trait, i.e., an organism resistant to that drug has been
found outside of a laboratory setting. Nonetheless, the consensus was
that the original language should be maintained. They noted that there
was no evidence that this section had failed to protect the public
health. They also noted that once resistance has occurred in the
microbial community outside of a laboratory setting, the use of such
strains in a contained laboratory environment poses no additional risk
to public health. Therefore, only those experiments that propose to
introduce resistance to a therapeutic drug, when such resistance does
not yet exist in the community, should require both RAC review and NIH
Director approval. As to whether a single documented case of drug
resistance is sufficient to allow this work to proceed without the
necessity of RAC review and NIH Director approval, at least one expert
noted that when there is a single case report, it is na[iuml]ve to
believe that there is only a single clinical isolate with that
resistance trait. There are probably dozens or hundreds of isolates
that were never reported and more that are undetected. The point is
that once resistance occurs naturally, as opposed to in a laboratory
setting, it is likely to occur again if acquisition of the antibiotic
resistance confers a survival advantage upon the organism.
The introduction of a drug resistance trait into organisms in a
laboratory setting when there are organisms outside the laboratory with
this same drug resistance trait is fundamentally different than
creating a novel drug resistant strain that does not exist outside of
the lab. While one expert commented initially that the focus should be
on resistance patterns in the U.S., others did not agree that such a
limited perspective was warranted. There was consensus that there
should be good documentation that this resistance exists outside of a
laboratory setting and a single case report may need to be confirmed.
Reports of clinical or environmental isolates should be the source of
documentation of resistance.
In sum, this section of the current NIH Guidelines appears to
protect public health adequately. There may indeed be some experiments
that raise important public health considerations but would not qualify
as Major Actions because there is a low level of documented resistance
to the drug that will be used for selection. However, it was not
possible to develop clear and easily interpretable criteria for
identifying such experiments. The solution proposed was to encourage
IBCs to consult with NIH/OBA and for NIH/OBA to consult with the RAC as
needed when there is an experiment that does not meet the criteria for
Section III-A-1-a but nonetheless raises important public health
questions.
There were very few comments on the proposed language regarding
analyzing subpopulations in determining the therapeutic usefulness of
any antibiotic. However, there was some concern that this language
might capture all antibiotics that could possibly be used rather than
being limited to those antibiotics that were used clinically.
Additional concern was raised about focusing on antibiotics that are
not commonly used in the U.S. and therefore whether the definition of
therapeutically useful should be limited to U.S. practice.
The intent of the proposed clarification regarding what is a
therapeutically useful drug was not meant to expand the requirement for
RAC review and NIH Director approval to all antimicrobials that might
exhibit in vitro activity against a microorganism, but rather to focus
on those that are used clinically as first or second line therapies in
certain populations. The additional language was intended to raise
awareness that the analysis of whether a drug is therapeutically useful
needs to include consideration of certain subpopulations, in particular
children and pregnant women, as many antibiotics may not be appropriate
for these specific populations. With respect to antibiotics not used in
the U.S., to the extent that certain pathogens have extensive impact on
international populations, it is prudent to consider the antibiotic of
choice in countries in which this pathogen causes disease. For example,
as background to the discussion of whether the transfer of
chloramphenicol resistance to Rickettsia typhi should be reviewed under
Section III-A-1-a, the investigators noted that chloramphenicol is
rarely used in the U.S. to treat disease caused by this organism.
However, as this disease has considerable impact worldwide, and in
particular in many developing countries in which chloramphenicol is
used, this antibiotic was considered to be a therapeutically useful
drug.
NIH/OBA agrees with the comments stating that the phrase ``not
known to acquire the trait naturally'' serves to identify the majority
of experiments that potentially pose higher risk to public health, and
therefore this language will be retained. One clarification to the
language was suggested by the RAC. Section III-A-1-a currently states
that the ``deliberate transfer of a drug resistance trait to
microorganisms that are not known to acquire the trait naturally, if
such acquisition could compromise the use of the drug to control
disease agents in humans, veterinary medicine, or agriculture, will be
reviewed by the RAC.'' As the introduction of a drug resistance trait
would normally eliminate that drug as a therapeutic option, the
analysis of whether this section applies has focused on whether the
acquisition of the resistance trait by that microorganism will
compromise the ability to control disease using alternative drugs.
Therefore, the wording has been clarified as follows:
[[Page 54595]]
The deliberate transfer of a drug resistance trait to
microorganisms that are not known to acquire the trait naturally (see
Section V-B, Footnotes and References of Sections I-IV), if such
acquisition could compromise the ability to control that disease agent
in humans, veterinary medicine, or agriculture, will be reviewed by the
RAC.
While there was consensus that this language adequately protected
public health for many years and served the scientific community, there
was acknowledgement that the mere fact that resistance to a drug has
been documented does not necessarily mean that there are no potential
public health concerns raised by use of that drug resistance trait in
that microorganism. These concerns may be handled by imposing
appropriate containment and other occupational health measures. In some
cases, an IBC may have adequate expertise from members with training in
infectious diseases to assess these risks and adopt appropriate
measures, but because other IBCs may not have that same expertise,
providing a mechanism for consultation with NIH/OBA or the RAC would be
helpful. In order to emphasize the fact that part of NIH/OBA's role is
to assist IBCs and other interested parties in evaluating containment
for recombinant and synthetic nucleic acid research, the following will
be added to Section III-A-1-a. This statement is a slight modification
to that found currently in Section IV-C-3 (Roles and Responsibilities
of the Office of Biotechnology Activities) of the NIH Guidelines.
At the request of an IBC, NIH/OBA will make a determination
regarding whether a specific experiment involving the deliberate
transfer of a drug resistance trait falls under Section III-A-1-a and
therefore requires RAC review and NIH Director approval. IBCs may also
consult with NIH/OBA regarding experiments that do not meet the
requirements of Section III-A-1-a but nonetheless raise important
public health issues. NIH/OBA will consult, as needed, with one or more
experts, which may include the RAC.
With respect to the comments about providing a list of drugs that
are clinically useful for a particular disease or to generate a list of
allowable transfers, inclusion of such information in the NIH
Guidelines is not appropriate. The drugs of choice for diseases are
often updated, and NIH/OBA follows the recommendation of the leading
medical textbooks and medical literature. Information on where to
obtain such guidance is already included in a Frequently Asked
Questions document on NIH/OBA's website under IBC Information http://oba.od.nih.gov/rdna_ibc/ibc.html. Experiments involving the deliberate
transfer of antibiotic resistance that present little or no risk to the
environment, agriculture, or public health, should be addressed in
informational guidances that are easily updated. Listing all acceptable
transfers of antibiotic resistance is not feasible.
Section III-A-1-a will now state:
The deliberate transfer of a drug resistance trait to
microorganisms that are not known to acquire the trait naturally
(see Section V-B, Footnotes and References of Sections I-IV), if
such acquisition could compromise the ability to control disease
agents in humans, veterinary medicine, or agriculture, will be
reviewed by the RAC.
Consideration should be given as to whether the drug resistance
trait to be used in the experiment would render that microorganism
resistant to the primary drug available to and/or indicated for
certain populations, for example children or pregnant women.
At the request of an Institutional Biosafety Committee, NIH/OBA
will make a determination regarding whether a specific experiment
involving the deliberate transfer of a drug resistance trait falls
under Section III-A-1-a and therefore requires RAC review and NIH
Director approval. An Institutional Biosafety Committee may also
consult with NIH/OBA regarding experiments that do not meet the
requirements of Section III-A-1-a but nonetheless raise important
public health issues. NIH/OBA will consult, as needed, with one or
more experts, which may include the RAC.
Section III-B. Experiments That Require NIH/OBA and Institutional
Biosafety Committee Approval
Once a Section III-A-I-a experiment is reviewed by the RAC and
approved by the NIH Director, equivalent experiments may not need to
follow the same approval process to determine the appropriate biosafety
containment level for the work. A new section under Section III-B
(Experiments that Require NIH/OBA and IBC Approval before Initiation)
was proposed to allow NIH/OBA (rather than the NIH Director) to review
and approve certain experiments deemed equivalent to those already
approved by the NIH Director, providing there is no new information
that would raise new biosafety or public health issues.
The following section is proposed to be added to the NIH
Guidelines:
Section III-B-2. Experiments that have been Approved (under
Section III-A-1-a) as Major Actions under the NIH Guidelines
Upon receipt and review of an application from the investigator,
NIH/OBA may determine that a proposed experiment is equivalent to an
experiment that has previously been approved by the NIH Director as
a Major Action, including experiments approved prior to
implementation of these changes. An experiment will only be
considered equivalent if, as determined by NIH/OBA, there are no
substantive differences and pertinent information has not emerged
since submission of the initial III-A-1 experiment that would change
the biosafety and public health considerations for the proposed
experiments. If such a determination is made by NIH/OBA, these
experiments will not require review and approval under Section III-
A.
Summary of Revised Language
The following provides the new language for the amended sections
discussed above.
Title of the NIH Guidelines
NIH Guidelines for Research Involving Recombinant or Synthetic
Nucleic Acid Molecules
Section I. Scope of the NIH Guidelines
Section I-A. Purpose
The purpose of the NIH Guidelines is to specify the practices
for constructing and handling: (i) recombinant nucleic acid
molecules, (ii) synthetic nucleic acid molecules, including those
that are chemically or otherwise modified but can base pair with
naturally occurring nucleic acid molecules, and (iii) cells,
organisms, and viruses containing such molecules.
Section I-B. Definition of Recombinant and Synthetic Nucleic
AcidsIn the context of the NIH Guidelines, recombinant and synthetic
nucleic acids are defined as:
(i) Molecules that a) are constructed by joining nucleic acid
molecules and b) can replicate in a living cell, i.e. , recombinant
nucleic acids;
(ii) Nucleic acid molecules that are chemically or by other
means synthesized or amplified, including those that are chemically
or otherwise modified but can base pair with naturally occurring
nucleic acid molecules, i.e., synthetic nucleic acids; or
(iii) Molecules that result from the replication of those
described in (i) or (ii) above.
Section I-C. General Applicability
Section I-C-1. The NIH Guidelines are applicable to:
Section I-C-1-a. All recombinant or synthetic nucleic acid
research within the United States (U.S.) or its territories that is
within the category of research described in either Section I-C-1-a-
(1) or Section I-C-1-a-(2).
Section I-C-1-a-(1). Research that is conducted at, or sponsored
by, an institution that receives any support for recombinant or
synthetic nucleic acid research from NIH, including research
performed directly by NIH.
An individual who receives support for research involving
recombinant or synthetic nucleic acids must be associated with or
sponsored by an institution that assumes the responsibilities
assigned in the NIH Guidelines.
[[Page 54596]]
Section I-C-1-a-(2). Research that involves testing in humans of
materials containing recombinant or synthetic nucleic acids
developed with NIH funds, if the institution that developed those
materials sponsors or participates in those projects. Participation
includes research collaboration or contractual agreements, not mere
provision of research materials.
Section I-C-1-b. All recombinant or synthetic nucleic acid
research performed abroad that is within the category of research
described in either Section I-C-1-b-(1) or Section I-C-1-b-(2).
Section I-C-1-b-(1). Research supported by NIH funds.
Section I-C-1-b-(2). Research that involves testing in humans of
materials containing recombinant or synthetic nucleic acids
developed with NIH funds, if the institution that developed those
materials sponsors or participates in those projects. Participation
includes research collaboration or contractual agreements, not mere
provision of research materials.
Section II-A-3. Comprehensive Risk Assessment
In deciding on the appropriate containment for an experiment,
the first step is to assess the risk of the agent itself. Appendix
B, Classification of Human Etiologic Agents on the Basis of Hazard,
classifies agents into Risk Groups based on an assessment of their
ability to cause disease in humans and the available treatments for
such disease. Once the Risk Group of the agent is identified, this
should be followed by a thorough consideration of how the agent is
to be manipulated. Factors to be considered in determining the level
of containment include agent factors such as: virulence,
pathogenicity, infectious dose, environmental stability, route of
spread, communicability, operations, quantity, availability of
vaccine or treatment, and gene product effects such as toxicity,
physiological activity, and allergenicity. Any strain that is known
to be more hazardous than the parent (wild-type) strain should be
considered for handling at a higher containment level. Certain
attenuated strains or strains that have been demonstrated to have
irreversibly lost known virulence factors may qualify for a
reduction of the containment level compared to the Risk Group
assigned to the parent strain (see Section V-B, Footnotes and
References of Sections I-IV).
While the starting point for the risk assessment is based on the
identification of the Risk Group of the parent agent, as technology
moves forward, it may be possible to develop an organism containing
genetic sequences from multiple sources such that the parent agent
may not be obvious. In such cases, the risk assessment should
include at least two levels of analysis. The first involves a
consideration of the Risk Groups of the source(s) of the sequences
and the second involves an assessment of the functions that may be
encoded by these sequences (e.g., virulence or transmissibility). It
may be prudent to first consider the highest Risk Group
classification of all agents that are the source of sequences
included in the construct. Other factors to be considered include
the percentage of the genome contributed by each parent agent and
the predicted function or intended purpose of each contributing
sequence. The initial assumption should be that all sequences will
function as they did in the original host context.
The Principal Investigator and Institutional Biosafety Committee
must also be cognizant that the combination of certain sequences in
a new biological context may result in an organism whose risk
profile could be higher than that of the contributing organisms or
sequences. The synergistic function of these sequences may be one of
the key attributes to consider in deciding whether a higher
containment level is warranted, at least until further assessments
can be carried out. A new biosafety risk may occur with an organism
formed through combination of sequences from a number of organisms
or due to the synergistic effect of combining transgenes that
results in a new phenotype.
A final assessment of risk based on these considerations is then
used to set the appropriate containment conditions for the
experiment (see Section II-B, Containment). The appropriate
containment level may be equivalent to the Risk Group classification
of the agent or it may be raised or lowered as a result of the above
considerations. The Institutional Biosafety Committee must approve
the risk assessment and the biosafety containment level for
recombinant or synthetic nucleic acid experiments described in
Sections III-A, Experiments that Require Institutional Biosafety
Committee Approval, RAC Review, and NIH Director Approval Before
Initiation; III-B, Experiments that Require NIH/OBA and
Institutional Biosafety Committee Approval Before Initiation; III-C,
Experiments that Require Institutional Biosafety Committee and
Institutional Review Board Approvals and NIH/OBA Registration Before
Initiation; and III-D, Experiments that Require Institutional
Biosafety Committee Approval Before Initiation.
Section III-A-1. Major Actions under the NIH Guidelines
The deliberate transfer of a drug resistance trait to
microorganisms that are not known to acquire the trait naturally
(see Section V-B Footnotes and References of Sections I-IV), if such
acquisition could compromise the ability to control disease agents
in humans, veterinary medicine, or agriculture, will be reviewed by
the RAC.
Consideration should be given as to whether the drug resistance
trait to be used in the experiment would render that microorganism
resistant to the primary drug available to and/or indicated for
certain populations, for example children or pregnant women.
At the request of an Institutional Biosafety Committee, NIH/OBA
will make a determination regarding whether a specific experiment
involving the deliberate transfer of a drug resistance trait falls
under Section III-A-1-a and therefore requires RAC review and NIH
Director approval. An Institutional Biosafety Committee may also
consult with NIH/OBA regarding experiments that do not meet the
requirements of Section III-A-1-a but nonetheless raise important
public health issues. NIH/OBA will consult, as needed, with one or
more experts, which may include the RAC.
Section III-B-2. Experiments that have been Approved (under Section
III-A-1-a) as Major Actions under the NIH Guidelines
Upon receipt and review of an application from the investigator,
NIH/OBA may determine that a proposed experiment is equivalent to an
experiment that has previously been approved by the NIH Director as
a Major Action, including experiments approved prior to
implementation of these changes. An experiment will only be
considered equivalent if, as determined by NIH/OBA, there are no
substantive differences and pertinent information has not emerged
since submission of the initial III-A-1 experiment that would change
the biosafety and public health considerations for the proposed
experiments. If such a determination is made by NIH/OBA, these
experiments will not require review and approval under Section III-
A.
Section III-C-1.
Experiments Involving the Deliberate Transfer of Recombinant or
Synthetic Nucleic Acid Molecules, or DNA or RNA Derived from
Recombinant or Synthetic Nucleic Acid Molecules, into One or More
Human Research Participants
Human gene transfer is the deliberate transfer into human
research participants of either:
Recombinant nucleic acid molecules, or DNA or RNA derived from
recombinant nucleic acid molecules, or
Synthetic nucleic acid molecules, or DNA or RNA derived from
synthetic nucleic acid molecules, that meet any one of the following
criteria:
a. Contain more than 100 nucleotides; or
b. Possess biological properties that enable integration into
the genome (e.g., cis elements involved in integration); or
c. Have the potential to replicate in a cell; or
d. Can be translated or transcribed.
No research participant shall be enrolled (see definition of
enrollment in Section 1-E-7) until the RAC review process has been
completed (see Appendix M-I-B, RAC Review Requirements).
Section III-F. Exempt Experiments
The following recombinant or synthetic nucleic acid molecules
are exempt from the
and registration with the Institutional Biosafety Committee is not
required; however, other federal and state standards of biosafety may
still apply to such research (for example, the Centers for Disease
Control and Prevention (CDC)/NIH publication Biosafety in
Microbiological and Biomedical Laboratories).
Section III-F-1. Those synthetic nucleic acids that: (1) can
neither replicate nor generate nucleic acids that can replicate in
any living cell (e.g. , oligonucleotides or other synthetic nucleic
acids that do not contain an origin of replication or contain
elements known to interact with either DNA or RNA polymerase), and
(2) are not designed to integrate into DNA, and (3) do not produce a
toxin that is lethal for vertebrates at an
[[Page 54597]]
LD50 of less than 100 nanograms per kilogram body weight. If a
synthetic nucleic acid is deliberately transferred into one or more
human research participants and meets the criteria of Section III-C
it is not exempt under this Section.
Section III-F-2. Those that are not in organisms, cells, or
viruses and that have not been modified or manipulated (e.g.,
encapsulated into synthetic or natural vehicles) to render them
capable of penetrating cellular membranes.
Section III-F-3. Those that consist solely of the exact
recombinant or synthetic nucleic acid sequence from a single source
that exists contemporaneously in nature.
Section III-F-4. Those that consist entirely of nucleic acids
from a prokaryotic host, including its indigenous plasmids or
viruses when propagated only in that host (or a closely related
strain of the same species), or when transferred to another host by
well established physiological means.
Section III-F-5. Those that consist entirely of nucleic acids
from a eukaryotic host including its chloroplasts, mitochondria, or
plasmids (but excluding viruses) when propagated only in that host
(or a closely related strain of the same species).
Section III-F-6. Those that consist entirely of DNA segments
from different species that exchange DNA by known physiological
processes, though one or more of the segments may be a synthetic
equivalent. A list of such exchangers will be prepared and
periodically revised by the NIH Director with advice of the RAC
after appropriate notice and opportunity for public comment (see
Section IV-C-1-b-(1)-(c), Major Actions). See Appendices A-I through
A-VI, Exemptions under Section III-F-6-Sublists of Natural
Exchangers, for a list of natural exchangers that are exempt from
the NIH Guidelines.
Section III-F-7. Those genomic DNA molecules that have acquired
a transposable element, provided the transposable element does not
contain any recombinant and/or synthetic DNA.
Section III-F-8. Those that do not present a significant risk to
health or the environment (see Section IV-C-1-b-(1)-(c), Major
Actions), as determined by the NIH Director, with the advice of the
RAC, and following appropriate notice and opportunity for public
comment. See Appendix C, Exemptions under Section III-F-8 for other
classes of experiments which are exempt from the NIH Guidelines.
Section IV-A. Policy
The safe conduct of experiments involving recombinant or
synthetic nucleic acids depends on the individual conducting such
activities. The NIH Guidelines cannot anticipate every possible
situation. Motivation and good judgment are the key essentials to
protection of health and the environment. The NIH Guidelines are
intended to assist the institution, Institutional Biosafety
Committee, Biological Safety Officer, and the Principal Investigator
in determining safeguards that should be implemented. The NIH
Guidelines will never be complete or final since all experiments
involving recombinant or synthetic nucleic acid molecules cannot be
foreseen. The utilization of new genetic manipulation techniques may
enable work previously conducted using recombinant means to be
accomplished faster, more efficiently, or at larger scale. These
techniques have not yet yielded organisms that present safety
concerns that fall outside the current risk assessment framework
used for recombinant nucleic acid research. Nonetheless, an
appropriate risk assessment of experiments involving these
techniques must be conducted taking into account the way these
approaches may alter the risk assessment. As new techniques develop,
the NIH Guidelines should be periodically reviewed to determine
whether and how such research should be explicitly addressed.
It is the responsibility of the institution and those associated
with it to adhere to the intent of the NIH Guidelines as well as to
their specifics. Therefore, each institution (and the Institutional
Biosafety Committee acting on its behalf) is responsible for
ensuring that all research with recombinant or synthetic nucleic
acid molecules conducted at or sponsored by that institution is
conducted in compliance with the NIH Guidelines. The following roles
and responsibilities constitute an administrative framework in which
safety is an essential and integral part of research involving
recombinant or synthetic nucleic acid molecules. Further
clarifications and interpretations of roles and responsibilities
will be issued by NIH as necessary.
Dated: August 29, 2012.
Lawrence A. Tabak,
Deputy Director, National Institutes of Health.
[FR Doc. 2012-21849 Filed 9-4-12; 8:45 am]
BILLING CODE 4140-01-P