1. Gretchen Hofmeister, April 2017
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 Guidelines specify 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 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

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)
Raka Mitra (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
rDNA work that requires notification to the IBC simultaneous to initiation
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
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

11. Know the NIH Guidelines “Section III”
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

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

13. Experiments that require NIH/OBA and IBC approval before initiation:
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.

14. Experiments that require IBC approval before initiation:
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

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 ( )
fully reconstructed 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. # )
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 Safety at U. Penn
St. Olaf College Biosafety Information
Contact Carleton or St. Olaf IBC members