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Biosafety Containment: Adeno-Associated Virus

Read about appropriate research practices with Adeno-associated virus (AAV) and AAV vectors.

Adeno-associated virus (AAV) and AAV viral vectors are useful tools for gene delivery and gene therapy. The virus and viral vector require a helper virus for replication because they are naturally replication deficient. This feature increases the safety of the viral vector. However, it requires management of work practices to separate AAV from common helper viruses like adenoviruses, herpesviruses, and papillomaviruses to reduce co-infection and unintended replication of AAV.

Adeno-associated virus and AAV vectors may be handled at Biosafety Level 1 (BSL-1) and Animal Biosafety Level 1 (ABSL-1) when they are:

  • Made in the absence of helper virus (such as Adenovirus and Herpes virus)
  • Used in the absence of any Risk Group 2 material, including human primary and established cell lines.
  • Free of any hazardous transgenes (such as an oncogene, toxin, shRNA from tumor suppressors, etc.)

If AAV is used with helper virus, Risk Group 2 Materials, or hazardous transgenes, then the AAV should be handled at BSL-2 or other BSL designation as identified in the approved Biological Use Authorization.

Work practices with AAV at BSL-1:

  1. Do not manipulate AAV in a biological safety cabinet at the same time as Risk Group 2 materials.
  2. Decontaminate work areas before and after use.
  3. If experiments with AAV also include Risk Group 2 materials or hazardous transgenes, BSL‐2 and ABSL‐2 containment rules apply.
  4. Work in animals:
    • Do not manipulate AAV in a biological safety cabinet or vivarium equipment at the same time as Risk Group 2 materials, including adenovirus.
    • Disinfect work surfaces before and after use.
    • Waste from animal work is disposed of as biohazardous. Examples include bedding and lab materials in contact with AAV. Carcasses must be disposed of as pathological waste. 
    • Use of engineered sharps are encouraged.


  • Meier AF, Fraefel C, Seyffert M. The Interplay between Adeno-Associated Virus and its Helper Viruses. Viruses 2020 Jun 19;12(6):662. Doi: 10.3390/v12060662. PMID: 32575422; PMCID: PMC7354565.
  • Naso MF, Tomkowicz B, Perry WL 3rd, Strohl WR. Adeno-Associated Virus (AAV) as a Vector for Gene Therapy. BioDrugs. 2017 Aug;31(4):317-334. doi: 10.1007/s40259-017-0234-5. PMID: 28669112; PMCID: PMC5548848.
  • Ronzitti G, Gross DA, Mingozzi F. Human Immune Responses to Adeno-Associated Virus (AAV) Vectors. Front Immunol. 2020 Apr 17;11:670. doi: 10.3389/fimmu.2020.00670. PMID: 32362898; PMCID: PMC7181373.
Contact EH&S Biosafety.