Overview of the NIH Guidelines for
Research Involving Recombinant
DNA Molecules
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
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
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*
What if we don’t comply with the
NIH Guidelines?
Non-compliance can result in:
1. Suspension, Limitation, or Termination of
NIH financial assistance for
•
The non-compliant research project
AND
•
ALL other rDNA research at the institution
2. Imposition of NIH overview and approval for
all rDNA projects before they are started
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
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
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)
• Functions to review, approve, and oversee
projects in accordance with the responsibilities
defined in NIH Guidelines Section 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
Institutional Biosafety Committee
(IBC) Responsibilities
On behalf of the institution, the IBC is responsible for:
1. Reviewing rDNA research conducted at or sponsored by the
institution
2. Setting containment levels
3. Notifying PI of the results of the IBC’s review and approval
4. Reporting significant problems or violations with the NIH
guidelines to NIH/OBA
5. Developing emergency plans covering accidental spills and
personnel contamination resulting from rDNA research
Principal Investigator (PI) Responsibilities
1. 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)
2. Be adequately trained in good microbiological techniques
3. Adhere to IBC approved emergency plans for handling accidental
spills and personnel contamination
4. Comply with applicable shipping requirements for recombinant DNA
molecules.
Principal Investigator (PI) Responsibilities
5. 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)
6. Remain in communication with the IBC throughout the
duration of the project
7. 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
Principal Investigator (PI) Responsibilities
8. 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:
a) rDNA work requiring NIH/OBA and IBC approval before initiation
–
Registration document required
b) rDNA work that must be approved by the IBC before initiation
–
Registration document required
c) rDNA work that requires notification to the IBC simultaneous to initiation
–
Registration document required
d) 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
Haverford’s rDNA Registration Document
You will be required to complete a registration document that looks like this:
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
• 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”
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
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.
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
•
–
III-B-1: Formation of rDNA containing genes for toxin molecules that are lethal to vertebrates
•
•
–
If the resistance trait can could compromise the use of a clinically significant drug
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)
“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:
1.
2.
3.
4.
IBC approval
IRB approval
IRB approved informed consent documents
Curriculum vitae of the PI (no more than 2 pages in biographical sketch
format)
5. NIH grant number (if applicable)
“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.
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
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)
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
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)
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
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
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
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 :
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
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
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
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
• Shirly Mildiner-Earley, shirlym@ehrs.upenn.edu
• Contact the IBC at Haverford (see Provost’s
Office)