Environmental Health & Safety

Viral Vectors Requirements

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PURPOSE

The NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (NIH Guidelines) require that Principal Investigators (PIs) seek and obtain Institutional Biosafety Committee (IBC) approval for all viral vectors. As part of the approval process, PIs must submit to the IBC information required by the NIH Guidelines, including the proposed Biosafety Level.

To assist PIs, this document summarizes IBC requirements for viral vectors currently approved at the University. Note: this document does not apply to human studies.

PERSONNEL AFFECTED

University of Rochester personnel who generate or use viral vectors and their supervisors

Environmental Health and Safety staff who audit labs

DEFINITIONS

Biosafety Level (BSL) refers to a set of work practices, equipment and facility design appropriate for working with infectious agents. Specific requirements are in BS020 - UR Biosafety Level Requirements for BSL1, ABSL1, BSL2, ABSL2, BSL2+ or ABSL2+.
G form: IBC Grant or Project Registration Form, includes the project description, a list of the vectors to be used, and the proposed Biosafety Level(s). For viral vectors, the G form in conjunction with the VV form meets the requirements of the NIH Guidelines.
NIH Guidelines: NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules
Replication-defective or Replication-deficient: describes viral vectors that cannot make additional copies of themselves
Transduction: the process by which foreign nucleic acids(DNA, RNA) are introduced into a cell by a viral vector
UR IBC: University of Rochester Institutional Biosafety Committee, composed of faculty, staff, and community members; required by the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules
Viral Vector: Partial viruses that deliver foreign genes into cells
VV form: IBC Viral Vector form, one for each viral base, includes the source of the vectors, the plasmids to be used, production methods, and the nucleic acid inserts that will be expressed.

RESPONSIBILITIES

It is the responsibility of the Principal Investigator and the Laboratory Supervisor to obtain IBC approval prior to starting work with viral vectors.

It is the responsibility of each person working with viral vectors to use them as approved by the IBC.

It is the responsibility of Environmental Health and Safety to periodically audit labs to ensure personnel are working at the appropriate Biosafety Level.

PROCEDURES

Most vectors are designed to be replication-defective. However, since no design system is perfect, replication-competent virus may be present (unless assayed). Therefore, the Biosafety Level is generally the same as the base virus.
Special concern is applied to those vectors or modifications that could:

extend the vector's host range (e.g. VSV-G pseudotyping allows entry into all cell types), tissue tropism, or enhance environmental stability
result in a replication-competent virus (vector design; using a helper virus)
integrate the vector into a portion of the genome leading to insertional mutagenesis
integrate an oncogene (e.g. RAS) or silence a tumor suppressor (e.g. p53) in the genome of a person (including in an exposure event)
generate a gene drive or selfish genetic element if injected into a person (i.e. a higher chance of a gene being inherited than by Mendelian genetics) - e.g. cassettes encoding Cas9 and sgRNA into a cut site or located adjacent to one another in the genome
deliver a gene encoding a toxin with a low LD₅₀

Replication-competent vectors are evaluated on a case-by-case base.
See the Appendix for a table of viral vectors and BSLs currently approved by the IBC.
To obtain IBC approval, submit G and VV forms. For forms, additional instructions and upcoming meeting schedule, visit www.safety.rochester.edu/homepages/ibchome.html.

REFERENCES

Akbari OS et al., BIOSAFETY. Safeguarding gene drive experiments in the laboratory. Science, 2015 349(6251):927-9, DOI: 10.1126/science.aac7932

Alemany R et al. Blood clearance rates of adenovirus type 5 in mice. J Gen Virol. 2000 Nov;81(Pt 11):2605-9, DOI: 10.1099/0022-1317-81-11-2605 (< 2 minutes)

Biosafety Considerations for Research with Lentiviral Vectors (NIH Recombinant DNA Advisory Committee), 2006 https://osp.od.nih.gov/wp-content/uploads/2014/01/Lenti_Containment_Guidance_0.pdf

Baldo A et al. General considerations on the biosafety of virus-derived vectors used in gene therapy and vaccination, Curr Gene Ther. 2013 Dec;13(6):385-94, PMCID: PMC3905712

Cavazzana M et al. Gene Therapy for X-Linked Severe Combined Immunodeficiency: Where Do We Stand? Hum Gene Ther. 2016 Feb;27(2):108-16 DOI: 10.1089/hum.2015.137

Jorgensen JH et al. editor, Manual of Clinical Microbiology, 11^th edition, 2015, American Society for Microbiology, www.asmscience.org/content/book/10.1128/9781555817381 (only UR personnel can obtain full access to this text using this link)

Kaiser J. Panel urges limits on X-SCID trials. Science. 2005 Mar 11; 307(5715):1544-5, DOI: 10.1126/science.307.5715.1544a

Ledford H. CRIPSR, the disruptor, Nature, 2015 522(7554):20-4, DOI: 10.1038/522020a

NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules, April 2016, https://osp.od.nih.gov/biotechnology/nih-guidelines/

Osakada F, Callaway EM. Design and generation of recombinant rabies virus vectors. Nat Protoc. 2013 Aug;8(8):1583-601, DOI: 10.1038/nprot.2013.094

Quetglas et al. Alphavirus vectors for cancer therapy. Virus Research 2010, 153:179-196, DOI: 10.1016/j.virusres.2010.07.027 (Sindbis vectors)

Tang SB and Levy JA. Inactivation of HIV-1 by trypsin and its use in demonstrating specific virus infection of cells. J Virol Methods. 1991 Jun;33(1-2):39-46, PMID: 1682337

Tenenbaum, L. et al. Evaluation of risks related to the use of adeno-associated virus-based vectors, Curr Gene Ther. 2003 Dec;3(6):545-65, PMID: 14683451

Wang et al. HSV-1 amplicon vectors are a highly efficient gene delivery system for skeletal muscle myoblasts and myotubes. Am J Physiol Cell Physiol. 2000. 278: 619-626, DOI: 10.1152/ajpcell.2000.278.3.C619

Wickersham, I. R., et al. (2007). "Retrograde neuronal tracing with a deletion-mutant rabies virus." Nature Methods 4: 47-49, PMID: 17179932, DOI:10.1038/nmeth999

Young AM et al. Failure of translation of human adenovirus mRNA in murine cancer cells can be partially overcome by L4-100K expression in vitro and in vivo. Mol Ther. 2012 Sep;20(9):1676-88, DOI: 10.1038/mt.2012.116 (oncolytic adenoviruses lack activity in murine cells)

APPENDICES/FORMS

Vectors/Biosafety Levels currently approved by the IBC - pdf version - List of Current Approved Levels

REVISION HISTORY

Date	Revision No.	Description
11/13/2011	New	Adenoviral Vector Requirements
06/26/2018	1	Consolidate BS014 (Biosafety Precautions for Replication-Incompetent Human Lentiviruses), BS026 (Biosafety Precautions for Replication-Incompetent Human Feline Immunodeficiency Virus) and the IBC guidance document General Information about Mammalian Virus Vectors into this policy/procedure, delete all BSL information redundant to BS020 - UR Biosafety Level Requirements for BSL1, ABSL1, BSL2, ABSL2, BSL2+ or ABSL2+, harmonize with Lab Safety Training, update references, update appendix table
09/18/2018	2	Update AAV BSL per 8/22/2018 IBC meeting

QUESTIONS or COMMENTS?
Contact EH&S at (585) 275-3241 or e-mail EH&S Questions.

This page last updated 8/11/2019. Disclaimer.

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