Recombinant DNA or Synthetic Nucleic Acid Molecules (r/sNA)

University Requirements for Working with Recombinant and Synthetic Nucleic Acid Molecules

Purpose

To provide an overview of the requirements for working with recombinant and synthetic nucleic acid molecules at the University of Minnesota. Recombinant and synthetic nucleic acid molecules are defined as:

(i) Molecules that a) are constructed by joining nucleic acid molecules and b) that can replicate in a living cell, i.e., recombinant nucleic acids;

(ii) Nucleic acid molecules that are chemically or by other means synthesized or amplified, including those that are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules, i.e., synthetic nucleic acids, or

(iii) Molecules that result from the replication of those described in (i) or (ii) above.

The intent of the following requirements is to provide a safe work environment and to meet compliance requirements.

Risk Assessment and Standard Operating Procedures (SOP) Development

Risk Assessment:

  • Assess the risk of the agent itself (e.g., host, vector). See Appendix B of the NIH Guidelines for risk group level for the biological agents known to infect human as well as selected animal agents that may pose theoretical risks if inoculate into humans.
  • Assess the risk caused by the manipulation of the agent. Agent factors to be considered:
    • Virulence
    • Pathogenicity
    • Infectious dose
    • Environmental stability
    • Route of transmission
    • Communicability
    • Operations
    • Quantity
    • Availability of vaccine or treatment
  • Assess the risk associated with the gene(s) to be transferred. What's the nature of the gene(s) to be transferred? Does the transgene encode either a potentially oncogenic gene product or a toxin molecule?
    Note: It is not possible to completely ensure that a replication-deficient virus will not gain back genes to become replication-competent.
    • Viral vectors do not need to be replication-competent.
    • It is possible for a "gene of interest" to be acquired by viruses or cells not expected to be associated with the gene.
  • Based on the risk assessment, determine the required level of physical and biological containment (BSL 1, 2, 3) in accordance with NIH Guidelines for Research Involving Recombinant and Synthetic Nucleic Acid Molecules.

SOP Development

Keeping in mind the above Biosafety Level determination, select the appropriate microbiological practices and laboratory techniques to be used for your specific research. Incorporate this information in written lab-specific Standard Operating Procedures (SOPs).

- The Writing Standard Operating Procedures (SOP) fact sheet provides the general guidelines how to develop an SOP.

Institutional Biosafety Committee (IBC) Oversight:

  • All recombinant and synthetic nucleic acid work, including work exempt from NIH Guidelines, must be approved by the IBC before work is started. Approved protocols are effective for three years with annual review.
  • Submit to the IBC online via eProtocol. For prompt processing, fill out the form completely and provide any additional pertinent information the committee will need to assess the risks associated with the proposal and the proposed biosafety levels, e.g.:
    • If animals are to be used in the recombinant and synthetic nucleic acid work, include information regarding shedding of the material by the animal in order to assess the biosafety level for animal housing.
    • Describe the safety features built into constructs such as removing all regions of homology with the packaging virus to prevent recombination resulting in replication-competent viruses.
  • The IBC approval process includes review of Standard Operating Procedures and proposed laboratory and animal biosafety levels with regards to the associated inherent risks and the proposed measures to be taken to reduce the risks. Research may not commence until IBC approval is granted.

General Practices