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Projects Requiring Approval

All projects involving biological agents must be reviewed and approved by the Institutional Biosafety Committee (IBC) prior to commencement of the work (see Scope on page 1-1 for a listing of biological agents). Principal Investigators must submit a biological use protocol, also known as a “Memorandum of Understanding and Agreement” (MOUA) to the IBC in order to initiate the approval process. The MOUA form is available on the EH&S “forms and applications” page, under “Biological Safety Forms.”

Biological Agents

All work involving biological agents must be reviewed by the University of ÁùºÏ±¦µä, Reno Institutional Biosafety Committee (IBC) for adherence to NIH/CDC biosafety guidance published in the latest edition of , the latest edition of , applicable regulations, as well as University policies and current biosafety practice.

Biosafety Level 1 (BSL-1) and Animal Biosafety Level 1 (ABSL-1)

Organisms in this category are not known to cause disease in healthy human adults. IBC approval is required for all BSL-1 work. Principal Investigators must submit a MOUA to the IBC in order to initiate the approval process.

Biosafety Level 2 (BSL-2) or 3 (BSL-3) and Animal Biosafety Level 2 (ABSL-2) or 3 (ABSL-3)

All work involving biological agents classified as BSL-2 or BSL-3 must be reviewed by the IBC. Containment levels, facility requirements, and work practices will generally follow NIH/CDC guidance; however, the IBC can raise or lower these requirements as appropriate. Principal Investigators are required to submit a MOUA to the IBC in order to initiate the approval process.

Biosafety Level 4 (BSL-4) and Animal Biosafety Level 4 (ABS-L4)

Projects involving BSL-4 organisms are currently prohibited at University.

Recombinant and Synthetic Nucleic Acid

As a condition of funding from the National Institutes of Health (NIH), all research at the University involving recombinant or synthetic nucleic acid molecules must be conducted in accordance with the most current version of  (NIH Guidelines). PIs are required to make an initial determination of the required biological and physical containment required. The approval level required for the proposed research is dependent on the NIH category to which the work corresponds. Approval by the IBC is required for all proposed experiments involving recombinant or synthetic nucleic acid molecules and transgenic animals and plants, including those exempt from NIH Guidelines. Principal Investigators must submit a MOUA to the IBC in order to initiate a request for approval. The following paragraphs summarize experiments covered by the NIH Guidelines and the required level of approval. Refer directly to the NIH Guidelines for a more detailed description of experiments and specific requirements.

Experiments that Require NIH Director Approval and Institutional Biosafety Committee Approval Before Initiation (NIH Guidelines section III-A)

Experiments involving the deliberate transfer of a drug resistance trait to microorganisms that do not acquire the trait naturally, where such acquisition could compromise the use of the drug to control disease in humans, veterinary medicine, or agriculture are included in this category. These experiments are considered a “Major Action” and cannot be initiated without the submission of relevant information on the proposed experiment to the Office of Science Policy, National Institutes of Health (via e-mail to: NIHGuidelines@od.nih.gov) the publication of the proposal in the Federal Register for a minimum of 15 days of comment, and specific approval by NIH. Approval by the IBC is required prior to initiation of these experiments.

Experiments requiring IBC and NIH approval (NIH Guidelines section III-B)

Experiments in this category include the deliberate formation of recombinant or synthetic nucleic acid containing genes for the biosynthesis of toxic molecules with an LD50 for vertebrates less than 100 ng/kg. This includes microbial toxins such as botulinum toxins, tetanus toxins, and diphtheria toxin. NIH Office of Science Policy (OSP) and IBC approval is required prior to initiation of these experiments.

Experiments Involving Human Gene Transfer requiring IBC approval (NIH Guidelines section III-C)

These experiments involve the deliberate transfer of recombinant or synthetic nucleic acid, or DNA or RNA derived from recombinant or synthetic nucleic acid molecules, into human research participants (human gene transfer). Research cannot be initiated until the IBC and all other applicable institutional and regulatory authorizations and approvals have been obtained.

Experiments requiring IBC approval before initiation of work (NIH Guidelines section III-D)

This category includes the following categories of experiments:

1. The introduction of recombinant or synthetic nucleic acid molecules into Risk Group 2, 3, or 4 agents.
2. Cloning of DNA from Risk Group 2, 3, or 4 agents into nonpathogenic prokaryotic or eukaryotic host-vector systems.
3. The use of infectious DNA or RNA viruses or defective DNA or RNA viruses in the presence of helper virus in tissue culture systems.
4. The use of whole transgenic animals, including creation of transgenic animals, and viable recombinant or synthetic nucleic acid molecule-modified microorganisms tested on whole animals. Certain experiments involving whole transgenic animals are exempt from NIH Guidelines and do not require submittal of a MOUA. See the section below titled “Transgenic Animals” for additional information.
5. Genetic engineering of plants using recombinant or synthetic nucleic acid molecule methods, use of such genetically engineering plants for experimental purposes, or use of plants with microorganisms or insects that contain recombinant or synthetic nucleic acid molecules.
6. Culture of more than 10 liters of organisms that contain recombinant or synthetic nucleic acid molecules.
7. The use of influenza viruses generated by recombinant or synthetic methods,

Experiments requiring IBC notice simultaneous with initiation of work (NIH Guidelines section III-E)

Experiments in this category are low risk and can be conducted using BSL-1 containment. Examples include experiments involving components that are all derived from non-pathogenic prokaryotes or non-pathogenic lower eukaryotes, the formation of recombinant or synthetic nucleic acid molecules that do not contain more than two-thirds of the genome of any eukaryotic virus, and plant and animal work (including the generation of transgenic rodents) involving recombinant or synthetic nucleic acid molecule methods that can be conducted at BSL-1. IBC approval is required and a MOUA must be submitted simultaneous with initiation of the experiments.

NIH exempt experiments (NIH section III-F)

The experiments listed below are exempt from the NIH Guidelines; however, University biosafety policy requires the PI to submit a MOUA to the IBC simultaneous with initiation of the work, with subsequent approval by the IBC. Exceptions to the requirement to submit a MOUA for NIH exempt experiments involving transgenic animals are described in the section titled “Transgenic Animals.”

The following recombinant or synthetic nucleic acid molecules are exempt from the NIH Guidelines:

1. Synthetic nucleic acids that cannot replicate or generate nucleic acids that can replicate in any living cell, and not designed to integrate into DNA, and do not produce a toxin with an LD50 for vertebrates of less than 100 ng/kg bodyweight.
2. Recombinant or synthetic nucleic acid molecules that are not in organisms, cells, or viruses and that have not been modified or manipulated to make them capable of penetrating cellular membranes.
3. Recombinant or synthetic nucleic acid molecules that consist solely of the exact recombinant or synthetic nucleic acid sequence from a single source that exists contemporaneously in nature.
4. Recombinant or synthetic nucleic acid molecules that consist entirely of nucleic acids from a prokaryotic host including its indigenous plasmids or viruses when propagated only in that host or when transferred to another host by well-established physiological means.
5. Recombinant or synthetic nucleic acid molecules consisting entirely of nucleic acids from a eukaryotic host including its chloroplasts, mitochondria, or plasmids (excluding viruses) when propagated only in that host.
6. Recombinant or synthetic nucleic acid molecules consisting entirely of DNA segments from different species that exchange DNA by known physiological processes, though one or more of the segments may be a synthetic equivalent.
7. Genomic DNA molecules that have acquired a transposable element provided the transposable element does not contain any recombinant and/or synthetic DNA.
8. Recombinant or synthetic nucleic acid molecules that do not present a significant risk to health or the environment, as listed in the NIH Guidelines, Appendix C.

Transgenic Animals

The acquisition, creation, and breeding of transgenic animals, including knockout animals, is included in the scope of the NIH Guidelines, and therefore, requires submittal of a MOUA for IBC approval prior to initiation of the work, with additional information and exceptions to this requirement provided below.

Resources:

• – May 2019 can be helpful when working with transgenic animals

Experiments Involving Transgenic Rodents

The creation of transgenic rodents that can be housed under BSL-1 containment is covered under Section III-E-3 of the NIH Guidelines and requires submittal of a MOUA simultaneous with initiation of the work, with subsequent approval by the IBC. The creation of transgenic rodents that must be housed at BSL-2 or higher containment levels is covered under Section III-D-4 and requires submittal of a MOUA to the IBC, with approval by the IBC prior to initiation of the work.

Breeding Transgenic Rodents

Breeding of two different lines of transgenic rodents, including knockout lines, can potentially generate a new line of transgenic rodent and is therefore covered by the NIH Guidelines. The breeding of two different lines of transgenic rodents may require submittal of a MOUA for IBC approval. The following experiments involving breeding of transgenic rodents do not require submittal of a MOUA.

The breeding of a transgenic rodent and a non-transgenic rodent with the intent of creating a new strain of transgenic rodent that can be housed at BSL-1 containment will not require submittal of a MOUA if both parental rodents can be housed at BSL-1 containment and:

1. Neither parental transgenic rodent contains more than one-half of the genome of an exogenous eukaryotic virus from a single family of viruses, or a transgene that is under the control of a gammaretroviral long terminal repeat, and
2. The transgenic rodent that results from this breeding is not expected to contain more than one-half of an exogenous viral genome from a single family of viruses.

Breeding within the same line of transgenic rodents, including knockout lines, at BSL-1 is exempt from the NIH Guidelines and submittal of a MOUA to the IBC is not required. If the transgenic rodents require housing at BSL-2 or higher containment level, then the breeding is covered by section III-D-4-b and requires submittal of a MOUA and IBC approval before initiation.

Breeding of Transgenic Animals Other Than Rodents

The breeding of all other transgenic animals is covered by the NIH Guidelines section III-D-4-a or III-D-4-b, depending on the containment level required, and requires submittal of a MOUA and prior approval by the IBC.

Purchase or Transfer of Transgenic Animals

The purchase or transfer of transgenic rodents, including knockout rodents, which can be housed at BSL-1, is exempt from the NIH Guidelines and submittal of a MOUA is not required. The purchase or transfer of transgenic rodents that must be housed at BSL-2 or higher containment is covered by section III-D-4 and requires submittal of a MOUA with IBC approval before purchase or transfer. The purchase or transfer of any other animal other than rodent species, at any biosafety level, is covered by the NIH Guidelines and requires submittal of a MOUA and IBC approval prior to purchase or transfer.

MOUA Amendment and Termination

Changes involving new biological agents, significant procedural changes, or any other modifications must be approved by the IBC. PIs wanting to modify a current MOUA are required to submit a MOUA Amendment form to the University Biosafety Officer (The IBC is now using Safety Stratus to process MOUA and amendment, contact Keith Kikawa or Chet Carpenter if you have questions about how to submit a MOUA or an amendment). Significant amendments will require review by the IBC. The PI is required to notify the University Biosafety Officer when a project is no longer active and must also indicate that the biological agents associated with the research have either been destroyed/inactivated or transferred to another PI with and active MOUA through an amendment.

MOUA Expiration and Annual Update

MOUAs are approved for three years; however, PIs are required to verify and update approved projects on an annual basis. The purpose of the annual update is to allow the PI to verify continuance of the project, discontinue the project, or amend the MOUA. The University Biosafety Officer or other designated person will contact each PI and request verification of the accuracy of the MOUA. The PI is required to update any incorrect information by submitting a MOUA Amendment form. Significant modifications will require IBC approval.

Additional Information

1. NIH Principal Investigator Responsibilities Under the NIH Guidelines.
2. NIH Guidance on Biosafety Considerations for Research with Lentiviral Vectors.
3. NIH Genetically Modified (Transgenic) Animals and the Use of Recombinant or Synthetic Nucleic Acid Molecules in Animals.
4. FAQs for Research on Genetically Modified (Transgenic) Animals – May 2019 can be helpful when working with transgenic animals

Chapter 3: Biosafety Regulations and Guidelines