Overview of the NIH Guidelines for Research Involving Recombinant DNA Molecules This is an edited combination of Haverford College’s IBC training materials and training materials from the NIH; by Gretchen Hofmeister, April 2017

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 April 2016

Why does Carleton need to oversee rDNA research? Carleton receives NSF funding for non-exempt projects involving rDNA research, therefore: ALL researchers at Carleton 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

How can Carleton and St. Olaf 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

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)

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

Institutional Biosafety Committee (IBC) Functions to review, approve, and oversee projects in accordance with the responsibilities defined in NIH Guidelines Section IV-B-2 Committee membership requirements: Individuals with expertise in rDNA technology Individuals with expertise in human gene transfer* Individual with expertise in plant containment* Scientist with expertise in animal containment* Biosafety officer¶ Community members not affiliated with institution *when experiments involve plants, animals, or humans ¶For research at BL3 or BL4

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

NIH Guidelines – Section II Safety Considerations Risk assessments: (Appendix B) RG 1 RG 2 RG 3 RG 4 Agents that are not associated with disease in healthy adult humans Agents that are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available Agents that are associated with serious or lethal human disease for which preventive or therapeutic interventions may be available (high individual risk but low community risk) Agents that are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available (high individual risk and high community risk)

NIH Guidelines – Section II Safety Considerations Containment Physical (Appendix G) Practices Equipment Facilities Biological (Appendix I) Survival Transmission BSL4 BSL3 BSL2 BSL1

Section III - Levels of Review IBC, RAC, NIH Director IBC, OBA (in consult with experts) IBC, IRB, RAC IBC IBC (notification) Exempt RISK

NIH Guidelines - Section III Levels of Review Level of review Example of types of research covered Relevant section(s) of the NIH Guidelines IBC, RAC review, and NIH Director review and approval Experiments that compromise the control of disease agents in medicine through deliberate transfer of a drug resistance trait III-A IBC approval and NIH review for containment determinations Experiment involving the cloning of toxin molecules with LD50 of less than 100 nanograms per kilogram of body weight III-B IBC and IRB approval and NIH review before research participant enrollment Experiments involving the deliberate transfer of recombinant or synthetic nucleic acid molecules into a human research participant III-C IBC approval before initiation Creating stable germline alterations of an animal’s genome, or testing viable recombinant or synthetically modified microorganisms on whole animals, where BL-2 containment or greater is necessary III-D IBC notice at initiation Creating stable germline alterations of rodents by introduction of recombinant or synthetic nucleic acid molecules when these experiments require only BL-1 containment III-E Exempt from the NIH Guidelines. IBC registration not required if experiment not covered by Sections III-A, III-B, or III-C Purchase or transfer of transgenic rodents III-F

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

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

Carleton’s rDNA Registration Review Process Submit the completed and signed registration forms to the chair of the IBC (Associate Dean of the College) 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

Carleton’s 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 (if applicable) 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

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 Safety at U. Penn St. Olaf College Biosafety Information Contact Carleton or St. Olaf IBC members