On September 5th, 2012, the National Institutes of Health, Office of Biotechnology Activities, Office of Science Policy (NIH/OBA) published a Federal Register Notice entitled “Final Action Under the NIH Guidelines for Research Involving Recombinant DNA Molecules (NIH Guidelines)”. This Notice was to inform the public regarding final changes to the “NIH Guidelines for Research Involving Recombinant DNA Molecules”, as proposed in the notice 74 FCR 9411, published on March 4th, 2009. The proposed revisions were to (1) address biosafety considerations for synthetic nucleic acids, and (2) modify criteria regarding whether introduction of drug resistance into a microorganism must be reviewed by the Recombinant DNA Advisory Committee (RAC) and approved by the NIH Director. Public comments were taken and reviewed, and public meetings were held on June 23rd, 2009, June 16th, 2010, and June 16th, 2010 to discuss these topics.
Regarding the first proposed change, the committee approved a change in the title of the NIH Guidelines, from “NIH Guidelines for Research Involving Recombinant DNA Molecules” to “NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acids Molecules”. The committee decided that the since the majority synthetic DNA used in research is replication-incompetent, there is no need for the NIH to provide additional oversight.
Furthermore, there is new language in the Purpose of the NIH Guidelines, from:
“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.” 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 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”.
This change states what synthetic DNA is, and that the review of primers or other small DNAs that are not introduced into an organism will not need to be reviewed. The definition of Recombinant Molecules has been modified as well (Section I-B) to:
“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.”
Regarding recombinant DNA transfer to human subjects, RAC review must occur before enrollment of patients:
“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).”
The Exempt experiments Section (Section III-F) has been modified, including the introduction, to reflect that although the listed DNA does not need to be registered, there may be some biosafety implications:
“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).”
Other exemptions include:
Section III-F-2 exempts DNA not found in a host (organism, virus, cell) or been manipulated to enable penetration of cell membranes
Section III-F-3 exempts sequences that are exact copies of naturally occurring sequences
Section III-F-4 exempts plasmids or sequences in prokaryotes when propagated only in that host, or are transferred to a closely related host
Section III-F-5 exempts plasmids or sequences in eukaryotes when propagated only in that host, or are transferred to a closely related host. These include sequences found in mitochondria, chloroplasts, or naturally occurring plasmids.
Section III-F-6 exempts conjugation transferred sequences between known conjugators of one or more species (a list of conjugators is provided in an appendix)
Section III-F-7 exempts transposon-mediated genetic transfer, unless the transposon has been recombinantly modified
Section III-F-8 exempts all experiments not deemed to have “significant risk to health or environment” after a determination by the RAC and NIH Director. (Renumbering only, not a change in language or intent)
Another change is found in Section IV-A. Policy, where it is stated that the NIH Guidelines cannot predict and address every conceivable research possibility, so institutions and investigators should adhere to “the intent of the NIH Guidelines as well as to their specifics.”
Changes to Section II-A-3 (Comprehensive Risk Assessment) modify the required risk assessment (Risk Group) to include not only an analysis of the parent organism, but also take into account the nature and products of any recombinant or synthetic DNAs.
Section III-A-1 (Major Actions under the NIH Guidelines) where both the RAC and the NIH Director must approve the research includes modifying the language regarding introduction of DNA containing a factor that increases drug resistance. While proposed changes would have stated that no first line or second-line treatment resistances could be introduced, there are no changes to this section. An IBC may request a review by the NIH/OBA, on whether the research should be reviewed by the RAC and requires NIH Director approval.
Section III-B states that experiments equivalent to ones previously approved by the NIH Director can be approved by the IBC and OBA.
Proposed changes to Section III-E-1 of the NIH Guidelines regarding containment of recombinant DNA experiments when using two-thirds or less of a genome from certain viruses are still pending.
The new regulations will go into effect on March 5th, 2013. There is a six month lead time until then, to enable new research review procedures, and for outreach to PIs and investigators. All ongoing and proposed research that will be occurring on or after March 5th will need to be registered with the appropriate IBC. Further information can be found by contacting OBA by email or fax.