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February 12, 2025
by Resmi
Biosafety
Recombinant viruses are replication deficient and can infect a broad range of mammalian cell types. Please refer to your institutions Occuptation Health and Safety office for guidance on safe handling of viral particles. BrainVTA sends all viral particles on dry ice, If your virus did not arrive on dry ice, please contact us immediately.
Safety
● Adeno-Associated Virus (AAV)
Recombinant AAV constructs do not encode for either a potentially tumorigenic gene product or a toxin molecule. According to guidelines from the National Institutes of Health (NIH), recombinant AAV vectors can be handled in a Biosafety Level 1 (BSL-1). If dealing with biohazardous material then please handle under Biosafety Level 2 (BSL-2) containment.
● Lentivirus (LV)
Lentiviral vector constructs are derived from HIV and are therefore highly efficient vehicles for in vivo gene delivery. They can integrate transgenes into dividing and non-dividing cells, so work with lentiviral vectors must be carefully. NIH guidelines require the maintenance of a BSL-2 or BSL-2 enhanced control facility for work involving lentivirus.
Storage
● For short-term storage, all vectors could be stored at room temperature or 4°C.
● For long-term storage, keep all vectors at -80°C. Do NOT store at -20°C. Research suggests that long-term storage at -20°C may result in decreased transduction efficiency of the virus.
We recommend aliquoting your virus into desired volumes, snap-freezing in liquid nitrogen or a dry ice/ethanol bath before storing at -80°C for use in future experiments. Please avoid unnecessary freeze-thawing of the vectors, as excessive freeze-thawing cycles may result in a significant decrease in titer and loss of biological activity.
Handling
● Thawing prior to use
On ice or at room temperature, and use immediately.
● During experiment
Keeping on ice at all times.
● Cell Transduction
AAV particles from both small scale and large scale packaging can be applied directly for cell transduction.
Safety
● Adeno-Associated Virus (AAV)
Recombinant AAV constructs do not encode for either a potentially tumorigenic gene product or a toxin molecule. According to guidelines from the National Institutes of Health (NIH), recombinant AAV vectors can be handled in a Biosafety Level 1 (BSL-1). If dealing with biohazardous material then please handle under Biosafety Level 2 (BSL-2) containment.
● Lentivirus (LV)
Lentiviral vector constructs are derived from HIV and are therefore highly efficient vehicles for in vivo gene delivery. They can integrate transgenes into dividing and non-dividing cells, so work with lentiviral vectors must be carefully. NIH guidelines require the maintenance of a BSL-2 or BSL-2 enhanced control facility for work involving lentivirus.
Storage
● For short-term storage, all vectors could be stored at room temperature or 4°C.
● For long-term storage, keep all vectors at -80°C. Do NOT store at -20°C. Research suggests that long-term storage at -20°C may result in decreased transduction efficiency of the virus.
We recommend aliquoting your virus into desired volumes, snap-freezing in liquid nitrogen or a dry ice/ethanol bath before storing at -80°C for use in future experiments. Please avoid unnecessary freeze-thawing of the vectors, as excessive freeze-thawing cycles may result in a significant decrease in titer and loss of biological activity.
Handling
● Thawing prior to use
On ice or at room temperature, and use immediately.
● During experiment
Keeping on ice at all times.
● Cell Transduction
AAV particles from both small scale and large scale packaging can be applied directly for cell transduction.
FAQs
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What is recombinant AAV (rAAV)?Recombinant AAV is the artificial adeno-associated virus vector which does not contain any AAV rep and cap genes ( Rep and cap genes encode viral replication and capsid proteins, respectively). The gene of interest replace rep and cap and it is flanked by the ITRs which contain all the cis-acting elements necessary for replication and packaging.
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