Overview of the NIH Guidelines for Research Involving Recombinant DNA Molecules

1. Overview of the NIH Guidelines for Research Involving Recombinant DNA Molecules

2. What are the NIH Guidelines for Research Involving Recombinant DNA Molecules? • These guidelines specify practices for constructing and handling: • Recombinant DNA (rDNA) molecules • Organisms and viruses containing rDNA molecules • History: • Guidelines originally established in 1975 at the Asilomar Conference on Recombinant DNA • Established recommendations to perform rDNA experiments in a manner safe for researchers and the public • Revised as needed, most recently in Sept. 2009

3. What are rDNA molecules? • Definition of rDNA molecules: • Molecules constructed outside living cells by joining natural or synthetic DNA segments to DNA molecules that can replicate in a living cell • Synthetic DNA segments, likely to yield a potentially harmful polynucleotide or polypeptide, are considered equivalent to their natural DNA counterpart. • Examples: • Human gene transfer • Cloning plasmids • Viral or bacterial vectors • Transgenic animals • Genetically modified cell lines

4. Who must comply with the NIH Guidelines? • Guidelines are applicable to all rDNA research at any institution that receives NIH support for rDNA research. • This includes: • NIH-funded projects involving rDNA • Non-NIH funded projects involving rDNA • Research with HUMANS involving rDNA materials developed with NIH funds • Only if the institution sponsors,participates, or collaborates in any way to those projects • Not the mere provision of research materials *Individuals receiving NIH-funds for rDNA research must be associated with or sponsored by an institution*

5. What if we don’t comply with the NIH Guidelines? Non-compliance can result in: • Suspension, Limitation, or Termination of NIH financial assistance for • The non-compliant research project AND • ALL other rDNA research at the institution • Imposition of NIH overview and approval for all rDNA projects before they are started

6. What does all of this mean for Haverford researchers? Haverford receives NIH funding for projects involving rDNA research, therefore: • ALL researchers at Haverford must comply with the NIH Guidelines • Even those not directly receiving NIH funding. AND • Just one non-compliant researcher could jeopardize NIH grant funding for the entire college

7. How can Haverford stay in compliance with the NIH Guidelines? • The NIH Office of Biotechnology Activities (OBA)fosters awareness of and adherence to the NIH Guidelines • OBA periodically conducts site visits to review how institutions follow the NIH Guidelines • Compliance with the NIH Guidelines is the responsibility of: • The Institution • The Institutional Biosafety Committee (IBC) • The Principal Investigator

8. The Institution’s Responsibilities • Provide training tools to researchers on the safe use of rDNA • Inform Principal Investigators of their responsibilities outlined in the guidelines • Form an Institutional Biosafety Committee (IBC)

9. Institutional Biosafety Committee (IBC) • Functions to review, approve, and oversee projects in accordance with the responsibilities defined in NIH GuidelinesSection IV-B-2 • Committee membership requirements specified in Section IV-B-2 include: • Individuals with expertise in rDNA technology • Individuals with expertise in human gene transfer • Scientist with expertise in animal containment • Biosafety officer • Community members not affiliated with institution

10. Institutional Biosafety Committee (IBC) Responsibilities On behalf of the institution, the IBC is responsible for: • Reviewing rDNA research conducted at or sponsored by the institution • Setting containment levels • Notifying PI of the results of the IBC’s review and approval • Reporting significant problems or violations with the NIH guidelines to NIH/OBA • Developing emergency plans covering accidental spills and personnel contamination resulting from rDNA research

11. Principal Investigator (PI) Responsibilities • Make an initial determination of the required levels of physical and biological containment • Supervise the safety performance of the laboratory staff • Ensure the integrity of the physical containment (e.g. biological safety cabinets) • Ensure integrity of the biological specimens (e.g. purity and genotypic/phenotypic characteristics) • Be adequately trained in good microbiological techniques • Adhere to IBC approved emergency plans for handling accidental spills and personnel contamination • Comply with applicable shipping requirements for recombinant DNA molecules.

12. Principal Investigator (PI) Responsibilities • Submit an initial registration to the IBC for review and approval or disapproval • Report any subsequent changes to project (e.g. changes in the sources of DNA or host-vector system) • Remain in communication with the IBC throughout the duration of the project • Report to the IBC, NIH/OBA and other appropriate authorities: • Any significant problems, violations of the NIH Guidelines • Any significant research-related accidents and illnesses • Within 30 days of incident

13. Principal Investigator (PI) Responsibilities • On behalf of the institution, the PI is responsible for full compliance with the NIH guidelines in the conduct of rDNA research. PIs must be knowledgeable about the different types of rDNA work and their registration requirements: • rDNA work requiring NIH/OBA and IBC approval before initiation • Registration document required • rDNA work that must be approved by the IBC before initiation • Registration document required • rDNA work that requires notification to the IBC simultaneous to initiation • Registration document required • rDNA work that is exempt • While a registration document is NOT required by NIH, Haverford policy requires a record of all rDNA work No rDNA work requiring IBC review before initiation may be started or modified until full IBC approval is granted.

14. Haverford’s rDNA Registration Document You will be required to complete a registration document that looks like this:

15. It is the PI’s responsibility to know which section of the NIH Guidelines covers his or her rDNA work Penn’s registration form asks you to properly identify this information The following slides will help you determine appropriate categories Know the NIH Guidelines“Section III”

16. NIH Guidelines“Sections III-A, III-B, III-C” rDNA work requiring NIH/OBA and IBC approvalbefore initiation NIH Guidelines“Section III-D” rDNA work that must be approved by the IBC beforeinitiation NIH Guidelines“Section III-E” rDNA work that requires notification to the IBC simultaneous to initiation Know the NIH Guidelines“Section III”

17. NIH Guidelines“Sections III-A, III-B, III-C” covers: Experiments that require IBC and IRB and RAC review before research participant enrollment. Experiments involving the cloning of toxin molecules with LD50 of less than100ng/kg body weight. Experiments involving the deliberate transfer of rDNA, or DNA or RNA derived from rDNA, into one or more human research participants. Experiments that require NIH/OBA and IBC approval before initiation:

18. Examples of work under “Sections III-A, III-B, and III-C” Examples include but are not limited to: • III-A-1: Major actions requiring IBC, RAC, and NIH Director approval • III-A-1a: The deliberate transfer of drug resistance to microorganisms not naturally acquiring that resistance • If the resistance trait can could compromise the use of a clinically significant drug • III-B-1: Formation of rDNA containing genes for toxin molecules that are lethal to vertebrates • Toxins that have an LD50 of less than 100ng/kg body weight • Ex) Botulinum toxins, tetanus toxins, diphtheria toxin, Shigella dysenteriae neurotoxin • III-C-1: Experiments involving the deliberate transfer of rDNA into human research participants (human gene transfer)

19. “Section III-C-1”: Human Gene Transfer • Research involving the deliberate transfer of rDNA or DNA/RNA derived from rDNA into human research participants • NO research participants may be enrolled until the rDNA Advisory Committee(RAC) review has been completed • RAC review requirements outlined in “Appendix M” of the Guidelines • For clinical trial sites added after RAC review: • Clinical trial site must submit the following documents to NIH OBA: • IBC approval • IRB approval • IRB approved informed consent documents • Curriculum vitae of the PI (no more than 2 pages in biographical sketch format) • NIH grant number (if applicable)

20. “Section III-C-1”: Human Gene Transfer • NIH Guidelines apply if the institution: • Participates in project • Including research collaboration and contractual agreements • NOT merely providing research materials • PIs engaging in human gene transfer projects may delegate to another party the reporting functions set forth in “Appendix M” • This may be a corporate sponsor. • Written notification to NIH/OBA is required.

21. NIH Guidelines“Section III-D” covers: Experiments using risk group 2, 3 or 4 or restricted agents as host vector systems Experiments in which DNA from risk group 2, 3 or 4 or restricted agents is cloned into nonpathogenic prokaryotic or lower eukaryotic host-vector systems Experiments involving the use of infectious DNA or RNA viruses or defective DNA or RNA viruses in the presence of helper virus in tissue culture systems Experiments involving whole animals Experiments involving whole plants Experiments involving more than 10 liters of culture Experiments involving influenza virus Experiments that require IBC approval before initiation:

22. Examples of work under “Section III-D” Examples include but are not limited to: • III-D-1: Using lentiviral or adenoviral vectors • III-D-2: Gene inserts used are from pathogenic microorganisms • III-D-3: Helper virus is used in tissue culture to enhance pathogenicity of viral vectors • III-D-4: ANY rDNA materials that are going into animals • DOES NOT include making transgenic rodents • One of the most common categories of work at Penn • III-D-5: Genetically engineering plants using rDNA and/or using these plants in experiments • III-D-6: rDNA experiments using >10L of culture • III-D-7: rDNA work with all influenza viruses with special consideration of: • human H2N2 (1957-1968) • fully reconstructed 1918-1919 H1N1 • Highly Pathogenic Avian Influenza (HPAI) H5N1 (Goose/Guangdong/96-like lineage)

23. Experiments that require IBC notice simultaneous with initiation NIH Guidelines, “Section III-E” covers: • Only experiments that require BSL-1/ABSL-1 containment • If BSL-2/ABSL-2 containment is required, it cannot be “Section III-E” • rDNA molecules of eukaryotic viruses that contain no more than 2/3 of the viral genome • In tissue culture only and • Require BSL-1 containment • Must demonstrate the lack of helper virus for certain families of defective viruses being used • Experiments involving whole plants • Experiments involving transgenic rodents • Generation of transgenic rodents by altering genome through stable introduction of rDNA into germ-line

24. Examples of work under “Section III-E” • Examples include but are not limited to: • III-E-1: Recombinant eukaryotic virus containing < 2/3 of its genome is propagated in tissue culture • III-E-2: rDNA plant experiments requiring BSL-1 containment • III-E-3: Experiments with transgenic rodents requiring BSL-1 containment • Creating a transgenic mouse (BSL-1 containment)

25. NEW Exemption: Change to Section III-E-3 Requirements(as of January 19, 2011) Old Requirement: • Breeding of one or more transgenic rodents to create new transgenic rodents using BSL-1 conditions • Fell under section III-E-3 • Previously required registration

26. NEW Exemption: Change to Section III-E-3 Requirements(as of January 19, 2011) New Requirement: • Breeding of almost* all transgenic rodents that require BSL-1 housing is exempt • Exempt under section III-F-6 • No registration document required *Exceptions* The following must be registered: • Rodents that contain a transgene encoding >50% of an exogenous eukaryotic virus • Transgenic rodents in which the transgene is under control of a gammaretroviral promotor • Any breeding experiments that require BSL-2 conditions

27. Exempt rDNA Experiments • NIH Guidelines“Section III-F” covers exempt experiments • Registration with the IBC is not required • Exempt experiments are those involving rDNA molecules that: • III-F-1: are not in organisms or viruses • III-F-2: consist entirely of DNA from single nonchromosomal or viral DNA source • III-F-3: consist entirely of DNA from a prokaryotic host when propagated only in that host • III-F-4: consist entirely of DNA from an eukaryotic host when propagated only in that host (excluding DNA from viruses) • III-F-5: consist entirely of DNA from different species that exchange DNA by known physiological processes (list periodically updated in Appendices A-I through A-VI) • III-F-6: do not present significant risk to health or to the environment as determined by NIH Director with the advice of RAC

28. Dual Use Research • Definition: “Research that yields information or technologies with the potential to be misused to threaten public health or other aspects of national security.” • Identify this on Haverford’s rDNA registration form :

29. Training Requirements • All personnel who are listed on a registration document must complete training, including: • PI • Anyone directly involved in rDNA experiments • Registrations will not be approved until all training is complete • This module satisfies the knowledge portion of training on the NIH Guidelines • PI is responsible for “hands-on” training of personnel

30. Haverford’s rDNA Registration Review Process • Submit the completed and signed registration forms to the chair of the IBC (see Provost’s Office if you are unsure who is the current chair of the IBC - for 2013-2014,it is Rob Fairman) • A member of the IBC will review the registration form and contact you with any questions or necessary revisions • The IBC reviews registrations as needed • The IBC is not permitted to review registrations outside of a fully convened meeting SO • Please be sure to submit registrations in a timely fashion, allowing enough time for a full meeting of the IBC to be arranged and convened

31. Haverford’d rDNA Registration Review Process • If the IBC approves your registration, you will receive an approval letter with: • IBC registration number (ex. #11-805) • The biosafety level (BSL) required • The animal biosafety level (ABSL) required • Registrations must be renewed every 3 years • If your registration is for work requiring IBC approval BEFORE initiation (Sections III-A, III-B, III-C, III-D), you may not start work until an official approval letter has been received

32. Additional Resources to Foster NIH Guideline Understanding • Office of Biotechnology Activities website • •NIH Guidelines • Office of Environmental Health and Radiation Safety’s (EHRS) website for Biological Safetyat U. Penn • Shirly Mildiner-Earley, shirlym@ehrs.upenn.edu • Contact the IBC at Haverford (see Provost’s Office)