Overview of the NIH Guidelines for Research Involving Recombinant DNA Molecules

1. Overview of the NIH Guidelines for Research Involving Recombinant DNA Molecules This is an edited version of Haverford College’s IBC training materials Gretchen Hofmeister, April 2017

2. The NIH Guidelines for Research Involving Recombinant DNA Molecules • These Guidelinesspecify practices for constructing and handling: • Recombinant DNA (rDNA) molecules • Organisms and viruses containing rDNA molecules • The Guidelines are applicable to all research and teaching at Carleton that involving rDNA. • 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

3. 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)

4. 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: • 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

5. Institutional Biosafety Committee (IBC) • Gretchen Hofmeister (Associate Dean of the College), Chair of IBC • Chris Calderone(Carleton Assistant Professor of Chemistry) • Patrick Ceas(St. Olaf Chemical Hygiene Officer) • Shawn Galdeen(Carleton Biology Lab Manager) • RakaMitra(Carleton Associate Professor of Biology), Plant Expert • Jean Porterfield (St. Olaf Professor of Biology), Animal Expert • Wade Schulz (St. Olaf Biology Stockroom Manager) • Peggy Anne Hawkins (Veterinary Provisions, Inc./Aurora Pharmaceuticals Veterinarian, Director of Research, Microbiologist) • Dan Taylor (Northfield High School Science Teacher)

6. 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

7. 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 plans for managing spills at Carleton and St. Olaf. 4. Comply with applicable shipping requirements for recombinant DNA molecules.

8. 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

9. 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.

10. NIH Guidelines - Section III Levels of Review

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

12. 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”

13. 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:

14. 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:

15. 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 • 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)

16. 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

17. 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)

18. 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 Carleton’s rDNA registration form :

19. 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

20. 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

21. Carleton’s rDNA Registration Review Process • If the IBC approves your registration, you will receive an approval letter with: • IBC registration number (ex. #17-18-02) • 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

22. 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 • St. Olaf College Biosafety Information • Contact Carleton or St. Olaf IBC members