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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

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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:

Cloning plasmids
Viral or bacterial vectors
Transgenic animals
Genetically modified cell lines

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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

*Individuals receiving NIH-funds for rDNA research must be associated with or sponsored by an institution*

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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

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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

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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

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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)

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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
Scientist with expertise in animal containment
Biosafety officer
Community members not affiliated with institution

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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

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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.

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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

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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.

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Haverford’s rDNA Registration Document

You will be required to complete a registration document that looks like this:

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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

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Know the NIH Guidelines �“Section III”

NIH Guidelines “Sections III-A and III-B”

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

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

NIH Guidelines “Sections III-A and III-B” covers:

Experiments that require IBC and IRB 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.

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Examples of work under “Sections III-A and III-B”

Examples include but are not limited to:

III-A-1: Major actions requiring IBC 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

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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

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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)

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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

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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)

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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)

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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

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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

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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 :

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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

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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 2019-2020,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

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

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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-D), you may not start work until an official approval letter has been received

Haverford’d rDNA Registration Review Process

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Additional Resources to Foster NIH Guideline Understanding

Office of Biotechnology Activities website

•NIH Guidelines

Office of Environmental Health and Radiation SafetyOffice of Environmental Health and Radiation Safety’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)