Historically, the IBC has assigned all work with AAV/rAAV to Biosafety Level 2 (BSL-2) or Animal Biosafety Level 2 (ABSL-2). The following is guidance for determining the appropriate biosafety designation when working with AAV/rAAV vectors.

Is AAV a biohazard?

Historically, the IBC has assigned all work with AAV/rAAV to Biosafety Level 2 or Animal Biosafety Level 2 (BSL/ABSL-2). Although the NIH Guidelines does list AAV as a risk group one agent, it is under the CDC/OSHA regulations (Blood borne Pathogen Standard CFR 29: 1910- 1030) that the committee ruled.

Are AAV vectors safe?

So, the answer to my own question—are AAV vectors safe? —is an emphatic yes, as long as astronomically high doses are not administered, especially in older children and adults.

Is AAV hazardous?

Cultures of replication-defective AAV vectors are non-infectious and are not hazardous materials as defined by OSHA 1919.1200.

What is AAV serotype?

The recently identified AAV receptor (AAVR) is a key host receptor for multiple serotypes, including the most studied serotype, AAV2. AAVR binds directly to AAV2 particles and is rate limiting for viral transduction.

What is AAV payload?

Adeno-associated viral (AAV) vectors are an established and safe gene delivery tool to target the nervous system. However, the payload capacity of <4.9 kb limits the transfer of large or multiple genes. Oversized payloads could be delivered by fragmenting the transgenes into separate AAV capsids that are then mixed.

Why is AAV replication deficient?

Are recombinant AAVs replication deficient? For wild type AAV, replication has extremely low efficiency without the presence of helper virus, such as an adenovirus. For recombinant AAV produced nowadays, the replication and capsid genes are provided in trans (in pRep/Cap plasmid).

Is AAV safe for use in delivering recombinant genes into humans?

To date, AAV has been shown to be safe and effective in preclinical and clinical settings.

Is AAV gene therapy safe?

AAV is a natural virus that has been engineered into a delivery vector to carry therapeutic genes into the cells of patients with genetic diseases, such as haemophilia. AAV vectors are considered safe and usually do not insert their own genomes into the human genome.

How do you dispose of AAV?

SECTION V – METHOD OF DISPOSAL o Dispose of viral stock by autoclaving at 121°C for 30-45 minutes. Dispose of infected liquid cultures by decontamination with chlorine bleach (10% f.c.) for 10 minutes and then dispose of in sink.

Is lentivirus a biohazard?

Lentivirus is regarded as a biosafety level 2 material and safe to use due to its modified features (deletion of a number of accessory virulence genes , minimal genome of the viral particles, non-replicating and self-inactivation features), making it incapable of producing virus once infected into the host cell.

Do AAV vectors integrate?

AAV vectors can integrate by homologous recombination, which is termed AAV-mediated gene targeting [27]. In order to promote homologous recombination, a vector that contains homology arms derived from genomic chromosomal DNA that flank the genetic modification being introduced is used.

What is the biosafety level for adenovirus and AAV vectors?

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.

What is the biosafety level for recombinant AAVs?

Recombinant AAV constructs in which the transgene does not encode a potentially tumorigenic gene product or a toxin molecule and is produced in the absence of a helper virus can be handled in a Biosafety Level 1(BSL-1) facility. Otherwise, it should be handled as biohazardous material under Biosafety Level 2 (BSL-2) containment.

Can AAV be used in a biological safety cabinet?

Do not manipulate AAV in a biological safety cabinet at the same time as Risk Group 2 materials. Decontaminate work areas before and after use. If experiments with AAV also include Risk Group 2 materials or hazardous transgenes, BSL‐2 and ABSL‐2 containment rules apply.

What is the best storage temperature for AAV vectors?

Stability studies carried out in house and by colleagues show that purified AAV vectors are highly stable at temperatures of 4 C or lower. All viral vectors are shipped frozen on dry ice and should be stored at -80° C upon receipt and for long term storage. Vectors can be stored for short periods of time at -20 or +4°C.