[Federal Register: March 4, 2009 (Volume 74, Number 41)]
[Notices]
[Page 9411-9421]
From the Federal Register Online via GPO Access [wais.access.gpo.gov]
[DOCID:fr04mr09-56]
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DEPARTMENT OF HEALTH AND HUMAN SERVICES
National Institutes of Health
Office of Biotechnology Activities; Recombinant DNA Research:
Proposed Actions Under the NIH Guidelines for Research Involving
Recombinant DNA Molecules (NIH Guidelines)
AGENCY: National Institutes of Health (NIH), PHS, DHHS.
ACTION: Notice of consideration of a proposed action under the NIH
Guidelines.
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SUMMARY: In 2006, the National Science Advisory Board for Biosecurity,
an advisory committee to the Secretary of the Department of Health and
Human Services, the NIH Director and all Federal entities that conduct/
support life sciences research published a report
[[Page 9412]]
entitled ``Addressing Biosecurity Concerns Related to the Synthesis of
Select Agents.'' \1\ The report included a recommendation that the
United States Government (USG) ``examine the language and
implementation of current biosafety guidelines 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 USG adopted this
recommendation and asked NIH to review the NIH Guidelines for Research
with Recombinant DNA (NIH Guidelines) to evaluate whether these
guidelines need to be revised to address biosafety concerns for
research with synthetic DNA. With the advice of the NIH Recombinant DNA
Advisory Committee (RAC), which is responsible for advising the NIH
Director on all aspects of recombinant DNA technology, including
revisions to the NIH Guidelines, the following proposed changes were
developed. As outlined in more detail below, the proposed changes will
expand the scope of the NIH Guidelines to specifically cover nucleic
acid molecules made solely by synthetic means. The changes apply to
basic laboratory research and clinical research. In addition, changes
were made to clarify the criteria for determining whether an experiment
to introduce drug resistance into a microorganism raises important
public health issues such that it must be reviewed by the RAC and
approved by the NIH Director. Finally, the proposed amendments speak to
the appropriate level of review for recombinant or synthetic
experiments involving more than half but less than two-thirds of the
genome of certain viruses in tissue culture. These changes were
prompted by an increased understanding of the biology of certain
viruses that demonstrate there may be biosafety risks with certain
viruses that contain less than two-thirds of the viral genome.
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\1\ The full document is available at http://oba.od.nih.gov/
biosecurity/pdf/Final_NSABB_Report_on_Synthetic_Genomics.pdf.
DATES: The public is encouraged to submit written comments on this
proposed action. Comments may be submitted to OBA in paper or
electronic form at the OBA mailing, fax, and e-mail addresses shown
below under the heading FOR FURTHER INFORMATION CONTACT. All comments
should be submitted by May 4, 2009. All written comments received in
response to this notice will be available for public inspection in the
NIH OBA office, 6705 Rockledge Drive, Suite 750, MSC 7985, Bethesda, MD
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20892-7985, weekdays between the hours of 8:30 a.m. and 5 p.m.
FOR FURTHER INFORMATION CONTACT: If you have questions, or require
additional information about these proposed changes, please contact OBA
by e-mail at oba@od.nih.gov, or telephone at 301-496-9838. Comments can
be submitted to the same e-mail address or by fax to 301-496-9839 or
mail to the Office of Biotechnology Activities, National Institutes of
Health, 6705 Rockledge Drive, Suite 750, MSC 7985, Bethesda, Maryland
20892-7985. Background information may be obtained by contacting NIH
OBA by e-mail at oba@od.nih.gov.
SUPPLEMENTARY INFORMATION:
Background: 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 galvanized
segments of the scientific community. It also has captured the
attention of the general public and policymakers, prompting far-
reaching questions about the potential use of these techniques--
including the synthesis of novel forms of life. These techniques
promise to accelerate scientific discovery and have the potential to
yield new therapeutics for disease. This same technology may lead to
the modification of existing or the creation of new pathogens with
unexpected and potentially dangerous characteristics.
In 2004, the National Research Council (NRC) published a report
that made an important contribution to the development of biosecurity
policy for the biological sciences, ``Biotechnology in the Age of
Terrorism: Confronting the Dual Use Issue.'' \2\ While this report was
not the first to recognize this problem, and indeed the U.S. Government
(USG) had already initiated an examination of security issues in the
biological sciences, the NRC report laid out a series of actions to
improve biosecurity in life science research, one of which was the
creation of an advisory body. The USG recognized the need for such an
advisory body and formed the National Science Advisory Board for
Biosecurity (NSABB) to advise the U.S. Government on strategies for
minimizing the potential for misuse of information and technologies
from life sciences research, taking into consideration both national
security concerns and the needs of the research community. The NSABB,
as it is chartered, differs somewhat from the panel proposed by the NRC
report, but has aims similar to those envisioned by the NRC committee.
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\2\ The report is available from the National Academies Press:
http://www.nap.edu/catalog.php?record_id=10827#toc.
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At the NSABB's first meeting, the Secretary of Health and Human
Services tasked the NSABB with identifying potential biosecurity
concerns raised by the rapidly advancing ability to synthesize select
agents (7 CFR part 331, 9 CFR part 121, and 42 CFR part 73) and other
dangerous pathogens. In 2006, NSABB published a report entitled
``Addressing Biosecurity Concerns Related to the Synthesis of Select
Agents.'' \3\ 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 often educated in disciplines
that do not routinely include formal training in biosafety, e.g.,
engineering. These researchers may be uncertain about when to consult
an Institutional Biosafety Committee (IBC).
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\3\ The full document is available at http://oba.od.nih.gov/
biosecurity/pdf/Final_NSABB_Report_on_Synthetic_Genomics.pdf.
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The NSABB recommended to the Secretary of the Department of Health
and Human Services that the language and implementation of current
biosafety guidelines be examined to ensure that such guidelines and
regulation provide adequate guidance for working with synthetically
derived nucleic acids. This recommendation on the need for biosafety
guidance was considered by the Executive Branch through a trans-Federal
policy coordination process. The recommendation on the need for
biosafety guidance was accepted by the U.S. Government with the
understanding that implementation would be through modification of the
NIH Guidelines as appropriate. The changes to the NIH Guidelines would
then be cross-referenced in the Centers for Disease Control and
Prevention/NIH publication entitled: Biosafety in Microbiological and
Biomedical Laboratories (BMBL).
The Recombinant DNA Advisory Committee (RAC) considered the
applicability of the NIH Guidelines to the creation of, and experiments
with synthetic nucleic acids (``synthetic biology'') and whether the
NIH Guidelines adequately address the biosafety concerns that may arise
from this research. The proposed revisions to the NIH Guidelines are
intended to clarify the applicability of the NIH Guidelines to research
with synthetic nucleic acids and provide principles and procedures for
risk assessment and management of such research.
While the initial NSABB recommendation focused on synthetic
[[Page 9413]]
genomics, which is the synthesis of nucleic acids using chemical or
other methods that do not require traditional recombinant DNA
techniques, it was recognized that this may be only be the first step
in a research proposal. The synthetic nucleic acid will then likely be
placed in cells or organisms. As it is articulated in the NIH
Guidelines, it is the manipulation of the recombinant nucleic acids
that leads to different biosafety concerns. As such, the focus of any
review of synthetic genomics from a biosafety perspective needs to
address the biological experiments that will be carried out. Therefore,
with respect to the NIH Guidelines, the task was to review the
biosafety considerations of introducing these synthetic nucleic acids
into biological systems.
Synthetic genomics utilizes different techniques than traditional
recombinant methods of synthesis; however, the ultimate product may be
the same. The biosafety considerations in most cases are related to the
product being produced more than the technique used. In other words,
the technique for creating sequences of nucleic acids is not
determinative of virulence, transmissibility and pathogenicity of the
product, which are key considerations in biosafety. There is no one to
one correlation between increasing nucleic acid diversity and
increasing risk of harm. Indeed, what has developed in nature involves
complex and highly regulated sequences of nucleic acids in which there
is often synergy between genes. Bringing together a number of genes or
sequences from different sources may result in a nucleic acid sequence
that is not functional in an organism. On the other hand, a single
nucleic acid change which could be done by recombinant or synthetic
means could lead to a significant enhancement in virulence. The focus
of a biosafety analysis should be on the product with consideration of
the source of the sequences. Synthetic techniques may result in a
greater range of products than recombinant methods but the underlying
challenge is the same: trying to understand how those disparate parts
will act together. Ultimately a biological analysis of the end results
will be required.
Under the current risk assessment framework of the NIH Guidelines,
the starting point for any risk assessment begins with an assessment of
the parent organism from which the sequence is derived. As discussed
under Section II, Safety Considerations, synthetic techniques may
enable the synthesis of more complex chimeras containing sequences from
a number of different sources. This increasing complexity may make the
task of determining the parent organism more challenging. This is
addressed in proposed language that will be added to the risk
assessment section of the NIH Guidelines (see proposed changes to
Section II-A).
Therefore, the changes proposed below treat the biosafety risks of
experiments that use recombinant and synthetic techniques as
equivalent. Also, although it was recognized that synthetic genetic
manipulation techniques are not necessarily a very recent development,
the integration of other fields (for example, chemistry and
engineering) may lead to rapid development of yet unknown products that
may raise new biosafety risks not anticipated. The risk management
framework being presented herein is based on the current science and
that which appears to be feasible in the foreseeable future.
The amendments will broaden the scope of the NIH Guidelines, which
currently cover research involving DNA molecules created via
recombinant techniques (i.e., joining of DNA molecules), to encompass
nucleic acids that are synthesized chemically or by other means without
the use of recombinant technology. As amended, the NIH Guidelines will
apply to all nucleic acids. This is accomplished through changes in
Section I-A, Purpose and Section I-B, Definition of Recombinant DNA
Molecules. The required level of review will be based on the risk of
the experiment, i.e. the risk to the laboratory worker, the public and
the environment. Low risk basic research involving non-replicating
synthetic nucleic acids will be exempt from the NIH Guidelines and from
review at the local level. High risk basic and clinical studies may be
subject to review by the RAC and the NIH. To effect these changes, four
sections of the NIH Guidelines will be revised. The title of the
document will be changed to NIH Guidelines for Research Involving
Recombinant and Synthetic Nucleic Acid Molecules and throughout the NIH
Guidelines the term recombinant DNA will be changed to recombinant and
synthetic nucleic acids.
In addition to broadening the scope of the NIH Guidelines to
encompass synthetic nucleic acids, included are proposed amendments to
two other sections of the NIH Guidelines, Section III-A-1 and Section
III-E-1, in order to (1) clarify the oversight of recombinant
experiments involving the introduction of drug resistance traits and
(2) to change the level of review for recombinant or synthetic
experiments involving more than half but less than two-thirds of the
genome of certain viruses in tissue culture. These proposed amendments
were recommended by the RAC.
Section III-A-1 requires certain experiments involving the transfer
of drug resistance traits to microorganisms to be reviewed by the RAC
and approved by the NIH Director. The current language has raised
concerns from IBCs and investigators seeking to identify those
experiments that require this heightened review. The revisions to
Section III-A-1 will clarify that all experiments involving the
transfer of a drug resistance trait to a microorganism will be subject
to RAC review and NIH Director approval if the microorganism's
acquisition of the trait could compromise public health. The changes
will clarify that the microorganism's ability to acquire the trait
naturally is not relevant to the safety of the experiment, that the
provisions apply even if the drug at issue is not considered the ``drug
of choice,'' and that adverse effects on population subgroups need to
be considered.
Under the NIH Guidelines, approval for an experiment under Section
III-A is specific to the investigator submitting the proposal.
Recognizing that this may not be an efficient use of resources and may
slow important research, a new provision will authorize OBA to make a
determination that a proposed experiment that would fall under Section
III-A is equivalent to an experiment that has been reviewed previously
as a Major Action and approved by NIH Director. In such cases, OBA will
have the authority to permit this research to proceed without going
through RAC review and NIH Director approval if OBA determines that
there are no substantive differences in experimental design and
pertinent information has not emerged since submission of the initial
experiment that would impact on the biosafety or public health risks
for the proposed experiments.
Section III-E-1 of the NIH Guidelines currently states that tissue
culture experiments involving viral constructs that contain less than
two-thirds of the genome of any one of the high risk viruses may be
performed at the lowest containment level (Biosafety Level 1) and
initiated upon registration with the local institutional biosafety
committee. The change proposed to this section will increase the
threshold to less than one-half of the viral genome and require
evidence that the resulting nucleic acid molecules are not capable of
producing a replication competent virus. These changes are prompted by
an increased understanding of the biology of certain viruses for which
there may be biosafety
[[Page 9414]]
risks for research involving less than two-thirds of the viral genome.
These recommendations were adopted unanimously by the RAC at its
March 2008 meeting. Included in these proposed changes are targeted
questions that were considered in developing the proposed revisions to
the NIH Guidelines. NIH requests not only comments on the proposed
changes but also comment on the specific issues raised by these
questions.
It should be noted that the NIH Guidelines currently apply to
research that is conducted at or sponsored by institutions that receive
NIH funding for any research involving recombinant DNA. Due to these
proposed changes, the NIH Guidelines will apply to research that is
conducted at or sponsored by institutions that receive NIH funding for
any research involving recombinant DNA and synthetic acid molecules. In
addition, other, non-NIH, U.S. Government agencies, including the
Department of Defense, the Department of Veterans Affairs and
Department of 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 govern all Government funded research, it may be used as a tool
for the entire research community to understand the potential biosafety
implications of their research.
In reviewing the proposed changes it is important to understand
that NIH Guidelines outline appropriate biosafety practices and
containment measures for laboratory recombinant DNA (rDNA) research and
govern the conduct of clinical trials that involve the deliberate
transfer of rDNA, or DNA or RNA derived from rDNA, into human research
participants. The focus of the NIH Guidelines is on the risks to
laboratory workers, the public and the environment associated with rDNA
research and if implemented, synthetic nucleic acid research. The NIH
Guidelines do promote the use of biological containment through the
application of highly specific biological barriers that may limit the
infectivity, dissemination, or survival of recombinant agents outside
the laboratory. Biological containment may, therefore, mitigate the
consequences of intentional misuse of such agents but does not directly
address biosecurity issues raised by deliberate exposure outside of a
research setting. As revised, the NIH Guidelines will continue to focus
on the biosafety aspects of research with recombinant and synthetic
nucleic acid molecules.
There may also be biosecurity or dual use research concerns with
some research involving recombinant or synthetic nucleic acid
molecules, but that is beyond the scope of the NIH Guidelines.
Biosecurity aspects of research involving infectious agents are
addressed in other venues, including for example, in the CDC-NIH
Biosafety in Microbiological and Biomedical Laboratories, 5th Edition
(Section VI, Principles of Laboratory Biosecurity) and the Select Agent
Rules (42 CFR 73, 9 CFR part 121 and 7 CFR part 131). In addition, the
U.S.G. continues to address these issues. For example, the NSABB is
developing recommendations for the oversight of dual use research and
is also addressing the issue of personnel reliability among individuals
working with select agents.
Proposed Amendments to the NIH Guidelines
In order to ensure that biosafety considerations of synthetic
biology research are addressed appropriately, the NIH is proposing the
following changes to the NIH Guidelines:
Title of the NIH Guidelines
The title of the document is proposed to be changed from the NIH
Guidelines for Research Involving Recombinant DNA Molecules to the NIH
Guidelines for Research Involving Recombinant and Synthetic Nucleic
Acid Molecules.
Section I. Scope of the NIH Guidelines
In order to clarify the applicability of the NIH Guidelines to
research involving synthetic nucleic acids (NA), the following
modifications are proposed to Section I, Scope of the NIH Guidelines.
Section 1-A. Purpose
Section I-A (Purpose) of the NIH Guidelines currently states that:
``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 is proposed to be amended to read: ``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, or (iii) organisms
and viruses containing such molecules.''
As a result of these modifications, the NIH Guidelines will clearly
apply to both recombinant and synthetically derived nucleic acids,
including those that contain functional analogs of nucleotides (e.g. ,
those used in artificially engineered genetic systems).
In accordance with this change in the scope of the NIH Guidelines
the term ``recombinant DNA molecules'' will be replaced with
``recombinant and synthetic nucleic acid molecules.''
Section I-B. Definition of Recombinant and Synthetic Nucleic Acids
The current definition of recombinant DNA molecule in the NIH
Guidelines (Section I-B) is limited because it only explicitly refers
to DNA and requires that segments be joined, which may not need to
occur in research with synthetic NAs. The proposed revisions to the
definition would retain a definition of recombinant NA similar to the
current one for recombinant DNA but also add synthetic NA created
without joining of segments. The current definition of recombinant DNA
in Section I-B of the NIH Guidelines is articulated in three paragraphs
labeled as A, B, and C in this notice only. Paragraph A states: ``In
the context of the NIH Guidelines, recombinant DNA molecules are
defined as either: (i) Molecules that are constructed outside living
cells by joining natural or synthetic DNA segments to DNA molecules
that can replicate in a living cell, or (ii) molecules that result from
the replication of those described in (i) above.'' Paragraph B 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.'' Paragraph C 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 following modifications are proposed to Section I-B. Definition
of Recombinant DNA Molecules: Paragraph A is proposed to be revised to
read: ``In the context of the NIH Guidelines, recombinant and synthetic
nucleic acids are defined as: (i) Recombinant nucleic acid molecules
that are constructed by joining nucleic acid molecules and that can
replicate in a living cell, (ii) synthetic nucleic acid molecules that
are chemically, or by other means, synthesized or amplified nucleic
acid molecules that may wholly or partially contain functional
[[Page 9415]]
equivalents of nucleotides, or (iii) molecules that result from the
replication of those described in (i) or (ii) above.''
Paragraph B will no longer be included in the definition. It was
added to the NIH Guidelines in 1982 to clarify that then novel
synthetic DNA segments would be considered as equivalent to their
natural DNA counterparts with regards to containment conditions;
however, it only covered synthetic DNA if it produced a toxin or a
pharmacologically active agent. The language presented difficulty in
interpretation because of the lack of definition of ``toxin or a
pharmacologically active agent.'' Paragraph B is proposed to be deleted
due to the fact that the concepts are sufficiently covered in the
following portions: The new (ii) in paragraph A which explicitly
extends the scope of the NIH Guidelines to cover recombinant and
synthetic constructs, and Section III-F (Exempt Experiments) of the NIH
Guidelines, which as discussed later, exempts those synthetic nucleic
acid constructs that do not pose a significant biosafety risk.
Paragraph C will be deleted from this portion and will be moved to
Section III-F of the NIH Guidelines. This is a proposed reorganization
of the NIH Guidelines so that exempt molecules will be described in one
place. A new Section IIIF-7 is proposed to read: ``Genomic DNA
molecules of plants and bacteria that have acquired a transposable
element provided the transposable element does not contain any
recombinant or synthetic DNA'' are not subject to the NIH Guidelines.
In accordance with these changes in the scope and definition of the
NIH Guidelines, the term ``recombinant DNA molecules'' will be replaced
with ``recombinant and synthetic nucleic molecules'' throughout the NIH
Guidelines.
Section III-C-1. Experiments Involving the Transfer of Recombinant DNA,
or DNA or RNA Derived From Recombinant DNA, Into One or More Human
Research Participants
In accordance with the change to the scope and definition of
recombinant DNA, the definition of human gene transfer experiments will
be amended. The first paragraph of Section III-C-1 currently states:
``For an experiment involving the deliberate transfer of recombinant
DNA, or DNA or RNA derived from recombinant DNA, into 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).'' As amended the first paragraph will state: ``For an
experiment involving the deliberate transfer of recombinant or
synthetic nucleic acids into 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).''
Section III-F. Exempt Experiments
Additional modifications are proposed to augment or clarify
experiments that are exempt from the NIH Guidelines, those listed in
Section III-F. The exemptions under Section III-F are designed to
strike a balance between safety and overregulation. They exempt certain
nucleic acid molecules from oversight by the NIH Guidelines because
their introduction into a biological system is not expected to have a
biosafety risk that requires review by an IBC or the introduction of
these nucleic molecules into biological systems would be akin to
processes that already occur in nature and hence determining proper
biosafety practices would be evident by the characteristics of
naturally occurring sequence and/or would be covered by other
guidances. Is there a risk that these exemptions could inadvertently
exempt an experiment that is deserving of IBC review? First, it is
important to recognize that with the exception of the new proposed III-
F-1 discussed below, the exemptions from the original NIH Guidelines
have been preserved with minor modifications. While synthetic synthesis
of nucleic acids will potentially raise new biosafety concerns the
exemptions focus narrowly on a small set of products that should not
raise biosafety concerns that warrant IBC review whether created by
recombinant or synthetic means.
To emphasize that research exempt from the NIH Guidelines will
still have biosafety considerations and that other standards of
biosafety may apply, a modification is proposed to the introductory
language. 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 proposed to read: ``The following recombinant and/or synthetic
nucleic acids 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 CDC/NIH Biosafety in
Microbiological and Biomedical Laboratories Manual).''
Section III-F-1
A new exemption under Section III-F-1 will exempt synthetic nucleic
acids that cannot replicate from the NIH Guidelines unless they are
used in human gene transfer (see Section III-C-1). This exemption is
proposed so that the NIH Guidelines apply to synthetic NA 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 NA that can
not replicate or were derived from molecules that can replicate. The
biosafety risks of using such constructs in basic and preclinical
research are believed to be 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 identical to that observed with chemical
exposures, although nucleic acids are generally far less toxic than
most chemicals.
Members of the RAC Biosafety Working Group noted that one of the
original impetuses for creating a special biosafety oversight for
recombinant DNA research was the novel biosafety risks to the
individual laboratory worker, the public health, and the environment
presented by the ability of novel replicating nucleic acids to
disseminate and persist within and outside of the laboratory. This risk
of transmissibility is distinct from chemicals or other toxins, because
of the potential for long-term persistence.
Human gene transfer clinical trials should be differentiated from
basic research. Current human gene transfer trials often involve non-
replicating recombinant molecules. These are captured by the NIH
Guidelines (see Section III-C-1 and Appendix M), because they are
derived through recombinant technology that has steps involving
replication (e.g., replication incompetent vectors, RNAi or antisense
RNA expressed from vectors are all derived from replicating systems).
The biosafety and health risks for human gene transfer for synthetic
non-replicating nucleic acids are not fundamentally different from non-
replicating recombinant vectors.
The safety distinction between laboratory research and human gene
[[Page 9416]]
transfer is based on the difference in the potential health risk due to
inadvertent lab exposure during basic or preclinical work and
deliberate clinical gene transfer. The doses and routes of
administration used in human gene transfer generally increase the
risks. The 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, and immunological responses.
For example, in the context of human gene transfer, the deliberate
transfer of large numbers of replication incompetent retroviral vectors
to hematopoietic stem cells in human clinical trials for X-Linked
severe combined immunodeficiency disease contributed to the development
of leukemia in some subjects starting several years after dosing. This
is a unique situation in human trials that would not be replicated in a
preclinical lab setting. Human gene transfer also raises scientific,
medical, social and ethical considerations that warrant special
attention and public discussion.
The following new exemption is proposed to be inserted as Section
III-F-1; the current exemptions III-F-1 through III-F-5 are proposed to
be re-numbered as III-F-2 through III-F-6. Section III-F-6 is proposed
to become III-F-8, because a new section III-F-7 is proposed to be
inserted. Section III-F-1 is proposed to read:
Section III-F-1: Synthetic nucleic acids that can not replicate,
and that are not deliberately transferred into one or more human
research participants (see Section III-C and Appendix M).
In arriving at the conclusion that non-replicating synthetic
nucleic acids pose limited risks to the public or environment, the RAC
considered different types of potential experiments involving a range
of possible exposures (e.g., dose, route) and nucleic acids (e.g.,
positive strand RNA viruses, replication incompetent integrating
vectors). For most research, the risks were considered sufficiently low
so that little benefit was considered to be gained by increased
oversight, which may hinder research. However, some questions remained.
The public is encouraged to submit written comments on the following
questions raised by this proposed modification to distinguish between
laboratory and clinical research with replicating and non-replicating
NA molecules.
(1) Is there a sufficient distinction between the risks of basic
and preclinical research with replicating vs. non-replicating synthetic
molecules to warrant the exemption?
(a) What are the risks with the use of replication incompetent
integrating vectors in the laboratory? For example, preclinical
research with recombinant lentiviral vectors is covered by the current
NIH Guidelines because the vectors are generated using a step involving
replication. At the lower doses typically used in laboratory
experiments, are the risks to the laboratory worker of such non-
replicating, synthetic NA research sufficiently low as to warrant
exemption from the NIH Guidelines?
(2) Since the increased risk associated with human gene transfer is
in part related to the administration of higher doses, should the
exemption be limited to experiments involving the handling of low
quantities or doses of NAs? What quantity would not be expected to pose
a biosafety risk?
(3) Are there examples of non-replicating, synthetic NA research
that should not be exempt due to greater potential risks (e.g.,
expression cassettes for oncogenes or toxins)?
(4) For human gene transfer research, are there classes of non-
replicating molecules that should be exempt due to lower potential
risks (e.g., antisense RNA, RNAi, etc.)? If so, what criteria should be
applied to determine such classes?
Section III-F-2
Section III-F-1 is proposed to be renumbered to III-F-2 and will be
amended to clarify that replicating NAs that are not in cells (in
addition to organisms and viruses) are exempt. Essentially, 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 extremely unlikely to have biosafety risks.
The primary risks associated with all nucleic acids, whether
synthetic or natural, are the effects these can engender when inside an
organism or the cellular compartment. Nucleic acids can alter protein
expression patterns in cells by binding to nucleic acids and blocking
(1) replication of DNA, (2) transcription of DNA into RNA and (3)
translation of RNA into protein. Furthermore, binding of synthetic or
natural DNA to cellular nucleic acids may result in degradation of
cellular DNA or RNA through the activity of natural cellular defense
mechanisms. Natural or synthetic DNA may have catalytic activity (e.g.,
ribozymes) that can cleave target sequences in nucleic acids. It is
these effects that can potentially lead the cell or organism containing
the nucleic acid to pose a risk to laboratory workers, the public or
environment.
None of the effects described above will occur unless the nucleic
acid is introduced into an organism, or a cell. Nucleic acids, by
virtue of their physical and chemical properties do not readily
penetrate cell membranes. The negative charge of a nucleic acid
molecule effectively prevents transfer across the plasma membrane of a
cell unless the negative charges of the molecule are either masked or
neutralized by addition of chemical compounds (e.g., cationic lipids,
calcium phosphate) or the cell membrane is physically perforated (e.g.,
electroporation) to enable penetration and uptake by the cell.
In practice, the current NIH Guidelines cover the introduction or
modification of recombinant DNA in tissue culture, organisms and
viruses. Therefore, for clarity and in recognition that techniques have
developed to more readily permit introduction of nucleic acids into
cells, the amended F-1 speaks to cells, organisms and viruses. In
addition, as stated above, natural barriers exist for entry of
unmodified nucleic acids into cells. However, manipulation of molecules
modified for improved penetration of cell membranes in the laboratory
may have increased risk due to the enhanced ability to penetrate cell
membranes and thus be able to replicate. Therefore, section III-F-1 is
being modified to address such modified nucleic acids as well.
Specifically, Section III-F-1 is proposed to be renumbered as III-
F-2 and amended as follows:
The current Section III-F-1 states: ``Those that are not in
organisms or viruses.''
Section III-F-1 will be re-numbered to III-F-2 and is proposed to
be amended to: ``Section III-F-2. Recombinant or synthetic nucleic
acids 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.''
The proposed Sections III-F-3 through III-F-7 retain exemptions
that were in the original NIH Guidelines with minor revisions. In
reviewing these exemptions it is important to understand that it is not
the goal of the NIH Guidelines to regulate all nucleic acid research
but rather that subset of research that through recombinant or now
synthetic means results in unique organisms or cells that potentially
possess characteristics not yet seen in nature and hence pose potential
safety risks both to the individual as well as
[[Page 9417]]
the community should there be an inadvertent release. Specifically, the
molecules that fall under the new Section III-F-3 (formerly Section
III-F-2) are those that consist solely of the exact nucleic acid
sequence from a single source that exists contemporaneously in nature.
Those described in the new Sections F-4 and F-5 (formerly Sections F-3
and F-4) are nucleic acids that are being propagated in a host that is
either the natural host for such nucleic acids or is a closely related
prokaryotic or eukaryotic host. Again such constructs may already exist
outside of a laboratory. Research that falls under F-6 (formerly
Section F-5) is exempt because the manipulation of these nucleic acids
in a laboratory setting would be equivalent to that which occurs in
nature when certain organisms exchange genetic material via
physiological processes (e.g., bacterial mating) outside of a
laboratory setting. It is limited to those organisms that are already
known to exchange DNA in nature. Finally, research that falls under the
proposed Section F-7 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 in the NIH Guidelines of a recombinant DNA
molecule. Unless a transposon has been modified to be a recombinant
molecule, genomic DNA of either plants or bacteria that has acquired a
transposon is not subject to the NIH Guidelines. This is because if
these transposons have not been modified by the insertion of
recombinant or synthetic DNA, they are equivalent to what is already in
nature and the process occurs naturally outside of lab.
The following changes are proposed for the Section III-F
exemptions.
Section III-F-3
Section III-F-2 is proposed to be re-numbered to III-F-3 and
amended. In the current NIH Guidelines, research with molecules from a
single DNA source is exempt. This would include molecules containing
duplications or deletions; however, such molecules may present
different risks than those of the wild type parent agents. The revised
language is intended to clarify that exempt molecules must have the
exact nucleic acid sequence from an organism that currently exists in
nature in order to be exempt (e.g., because the 1918 influenza no
longer exists in nature, research involving the reconstructed virus
would not qualify for this exemption). The exemption does not imply
that there are no biosafety risks associated with such research but
rather recognizes that the NIH Guidelines do not apply to wild-type
strains currently found in nature because a risk assessment for such
work can be made with reference to the biological characteristics of
the wild-type organism and are covered by other NIH biosafety standards
(for example CDC/NIH Biosafety in Microbiological and Biomedical
Laboratories Manual).
The following modifications are proposed for Section III-F-2.
Section III-F-2 is proposed to be re-numbered to III-F-3 and amended as
follows:
The current III-F-2 states: ``Those that consist entirely of DNA
segments from a single nonchromosomal or viral DNA source, though one
or more of the segments may be a synthetic equivalent.'' III-F-2 is
proposed to be renumbered to III-F-3 and is proposed to be amended to
state: ``Recombinant or synthetic nucleic acids that consist solely of
the exact nucleic acid sequence from a single source that exists
contemporaneously in nature.''
This proposed modification would change ``single nonchromosomal or
viral source'' to simply ``single source.'' Specific comment is
requested as to whether it is sufficiently clear that single source
refers to ``single chromosomal, non-chromosomal, or viral NA source''
or should the language be specifically spelled out?
Section III-F-4
The current Section III-F-3 is proposed to be renumbered to Section
III-F-4 and amended. Section III-F-3 states: ``Those that consist
entirely of DNA 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.'' It is proposed to be
amended as follows: ``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
The current Section III-F-4 is proposed to be renumbered to Section
III-F-5. Section III-F-4 currently states: ``Those that consist
entirely of DNA 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).'' It is
proposed to state the following: ``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
The current Section III-F-5 is proposed to be renumbered to Section
III-F-6. The current Section III-F-5 states: ``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-5--Sublists of Natural Exchangers,
for a list of natural exchangers that are exempt from the NIH
Guidelines.'' It is proposed to be amended to state: ``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 A1-through A-VI will be
amended to reference Section III-F-6 rather than III-F-5.
Section III-F-7
A new Section III-F-7 is proposed to be added. This proposed new
Section takes an exemption that was previously included in the original
definition (Section I-B) and moves it to this Section so that the
definition of recombinant and nucleic acids found in the proposed
Section I-B is solely a definition and does not include exemptions. The
proposed exemption language has been simplified to make it clear that
unmodified transposons used in research are not subject to the NIH
[[Page 9418]]
Guidelines even if derived from a recombinant or synthetic system.
Section I-B: 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. New
Section III-F-7 is proposed to state:
Section III-F-7. Genomic DNA molecules of plants and bacteria
that have acquired a transposable element provided the transposable
element does not contain any recombinant or synthetic DNA.
Section III-F-8
The current Section III-F-6 is proposed to be renumbered to Section
III-F-8 and amended. This section provides a mechanism for the NIH
Director to expand the exemptions to molecules not covered elsewhere in
Section III-F. Research that falls under Section III-F-8 would need to
have been reviewed and approved by the NIH Director following advice
from the RAC and notice in the Federal Register to provide an
opportunity for public comment. Only research that has been deemed to
not present, following this extensive review process, a significant
risk to health or the environment would fall under this section.
Current Section III-F-6 states: ``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-6 for
other classes of experiments which are exempt from the NIH
Guidelines.'' Section III-F-6 is proposed to be amended to state:
``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 A1- through A-VI will be amended to reference
Section III-F-8 rather than III-F-6.
Section III-E-1. Experiments Involving the Formation of Recombinant DNA
Molecules Containing No More Than Two-Thirds of the Genome of Any
Eukaryotic Virus
Experiments covered by Section III-E-1 can be initiated using
Biosafety Level (BL) 1 containment simultaneously with Institutional
Biosafety Committee notice. Section III-E-1 currently states:
``Recombinant DNA molecules containing no more than two-thirds of the
genome of any eukaryotic virus (all viruses from a single Family being
considered identical [see Section V-J Footnotes and References of
Sections I-IV ]) may be propagated and maintained in cells in tissue
culture using BL1 containment. For such experiments, it must be
demonstrated that the cells lack helper virus for the specific Families
of defective viruses being used. If helper virus is present, procedures
specified under Section III-D-3, Experiments Involving the Use of
Infectious Animal or Plant DNA or RNA viruses or Defective Animal or
Plant DNA or RNA viruses in the Presence of Helper Virus in Tissue
Culture Systems, should be used. The DNA may contain fragments of the
genome of viruses from more than one Family but each fragment shall be
less than two-thirds of a genome.''
This section applies to viral constructs containing less than \2/3\
of the genome of any virus (with all viruses from a single Family being
considered as identical). However, concerns were raised that this level
of oversight may not be adequate for research with potential synthetic
biology agents derived from multiple segments of NA from a Family of
viruses. In addition, some wild type viruses (e.g., herpes viruses) may
be functional with less than \2/3\ of the genome present. Therefore,
the decision was made to propose to change \2/3\ to one-half of the
genome to reflect the current understanding of the biology of certain
viruses. While the use of a quantitative measure to define properties
of biological organisms is imperfect, the more conservative standard is
consistent with Appendix C-1 Recombinant DNA in Tissue Culture which
exempts from the NIH Guidelines recombinant DNA molecules from Risk
Groups 1 and 2 that contain less than one-half of any eukaryotic viral
genome. With this revision, experiments involving risk Group 3 and 4
viruses with less than one-half of any eukaryotic viral genome can be
initiated at BL1 containment simultaneously with IBC registration
provided evidence is also submitted attesting that the preparation(s)
are free of replication competent virus, which may be generated through
homologous recombination with endogenous proviruses or the use of a
helper virus. If revised as proposed, an investigator will be permitted
to initiate an experiment simultaneously with registration, since the
retention of a quantitative standard provides such clear guidance.
Section III-E-1 is proposed to be amended to state: ``Recombinant
and synthetic nucleic acid molecules containing no more than half of
the genome of any one Risk Group 3 or 4 eukaryotic virus (all viruses
from a single Family being considered identical [see Section V-J,
Footnotes and References of Sections I-IV ]) may be propagated and
maintained in cells in tissue culture using BL1 containment (as defined
in Appendix G) provided there is evidence that the resulting nucleic
acid in these cells are not capable of producing a replication
competent nucleic acid. For such experiments, it must be demonstrated
that the cells lack helper virus for the specific Families of defective
viruses being used. If helper virus is present, procedures specified
under Section III-D-3, Experiments Involving the Use of Infectious
Animal or Plant DNA or RNA viruses or Defective Animal or Plant DNA or
RNA viruses in the Presence of Helper Virus in Tissue Culture Systems
should be used. The nucleic acids may contain fragments of the genome
of viruses from more than one Family but each fragment shall be less
than one-half of a genome.''
Section IV-A Policy
Section IV-A concerns the roles and responsibilities of the local
institutions and investigators in implementing the NIH Guidelines. It
contains a general policy statement that is often evoked as the
``spirit'' of the NIH Guidelines because it acknowledges the inability
of the document to describe specifically all conceivable research or
emerging techniques; however, it remains the responsibility of
researchers and institutions to adhere to ``the intent of the NIH
Guidelines as well as to their specifics.'' In order to emphasize that
the NIH Guidelines are an evolving document which are expected to be
modified to address new developments in research or scientific
techniques, the following modifications are proposed to Section IV-A
(Policy).
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
[[Page 9419]]
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 proposed to be amended to read: ``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 experiments involving
recombinant and/or synthetic nucleic acids cannot be foreseen. The
utilization of new genetic manipulation techniques may enable work
previously done by 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 DNA 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. In addition, as the field
develops, new techniques and applications need to be monitored and
assessed to determine whether revisions to the NIH Guidelines are
needed. 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 recombinant and/or synthetic nucleic
acids 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 and/or synthetic nucleic acid molecules. Further
clarifications and interpretations of roles and responsibilities will
be issued by NIH as necessary.''
Section II. Safety Considerations
Currently, the risk assessment framework of the NIH Guidelines uses
the risk group of the parent organism as a starting point for
determining the necessary containment level. For example, genetic
modifications using 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 BL3 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 is applicable to experiments with synthetic nucleic
acids. However, additional language is proposed to provide further
guidance for evaluating research utilizing the capabilities of
synthetic biology, as use of these techniques may lead to the creation
of complex organisms for which identification of a parent organism, the
starting point of the existing recombinant DNA risk assessment, is more
difficult. Risk assessment may also be complicated by the limitations
in predicting function from sequence(s) or the synergistic effects from
combining sequences from different sources in a novel context.
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.
[[Page 9420]]
Individuals working with human immunodeficiency virus (HIV),
hepatitis B virus (HBV) or other bloodborne pathogens should consult
the applicable Occupational Safety and Health Administration
regulation, 29 CFR 1910.1030, and OSHA publication 3127 (1996 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 three paragraphs are proposed to be amended by inserting
the following two new paragraphs between the current first and second
paragraphs of Section II-A-3:
``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).
While the initial 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 a chimera in which the parent agent may not be
obvious. In such cases, the risk assessment should involve 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 an analysis of
the functional attributes of these sequences (e.g., sequence associated
with virulence factors, transmissibility, etc.). It may be prudent to
first consider the highest risk group classification of any agent
sequence included in the chimera. Other factors to be considered
include the percentage of the genome contributed by each of multiple
parent agents, and the predicted function or intended purpose of each
contributing sequence. The initial assumption should be that all
sequences will function as predicted in the original host context.
The IBC must also be cognizant that the combination of certain
sequences 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. A
new biosafety risk may occur with a chimera 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 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.''
Section III-A-1. Major Actions Under the NIH Guidelines
In reviewing the biosafety risks for synthetic genomics and biology
and the different levels of review for each experiment, the RAC
determined that it is important to also evaluate the class of
experiments that require the highest level of review. In doing so, it
was determined that the language for Section III-A-1 of the NIH
Guidelines (research involving the introduction of drug resistance)
does not clearly articulate the types of 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 multi-drug
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.''
Section III-A-1-a is proposed to be amended to: ``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 (see Section V-B, Footnotes and References of Sections
I-IV). 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.''
The deletion of the phrase ``that are not known to acquire the
trait naturally'' is proposed because the mechanism of acquisition
should not be relevant as to whether these experiments pose potential
public health risk and as such
[[Page 9421]]
should receive a higher level of review. Moreover, all forms of
antibiotic resistance occur naturally and the use of antibiotics
creates selective pressure for resistant strains. The additional text
recognizes that a drug may remain useful for control of a disease
despite some percentage of the population of microorganisms having
developed resistance. It is also intended to clarify that even if a
particular drug is not considered the ``drug of choice'' to treat a
disease, elimination of such a drug as a treatment option may still
raise important clinical and public health considerations for certain
subpopulations.
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 III-B (Experiments
that Require NIH/OBA and IBC Approval before Initiation) is proposed to
be added to allow NIH/OBA the discretion to review and approve certain
experiments if NIH/OBA determines that an equivalent experiment has
already been approved by the NIH Director and there are no substantial
changes to the proposed experiment or new information that would raise
new biosafety or public health issues. Under this proposal,
Investigators will be notified by NIH/OBA if such a determination has
been made.
The following addition is proposed to be added to Section III-B of
the NIH Guidelines to allow NIH/OBA the discretion to review and
approve certain experiments that have been previously reviewed by the
RAC and approved by the NIH Director as a Major Action.
``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 in
experimental design or pertinent information has not emerged since
submission of the initial III-A-1 experiment that would impact on the
biosafety or public health risks 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.''
Dated: February 26, 2009.
Amy P. Patterson,
Acting Director, Office of Science Policy, National Institutes of
Health.
[FR Doc. E9-4618 Filed 3-3-09; 8:45 am]
BILLING CODE 4140-01-P