Purpose

Ready-to-use AAV2 particles produced from pAAV-hSyn-DIO-mCherry (#50459). In addition to the viral particles, you will also receive purified pAAV-hSyn-DIO-mCherry plasmid DNA.

Delivery

• Volume100 µL
• Titer≥ 4×10¹² vg/mL
• Pricing$375 USD for preparation of 100 µL virus + $30 USD for plasmid.
• StorageStore at -80℃. Thaw just before use and keep on ice.
• ShipmentViral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.

Viral Production & Use

• Packaging Plasmidsencode adenoviral helper sequences and AAV rep gene, AAV2 cap gene
• BufferPBS + 0.001% Poloxamer 188
• SerotypeAAV2
• PurificationCsCl gradient ultracentrifugation
• Reporter GenemCherry (Cre-dependent)

Biosafety

Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. 

Addgene Comments

Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.1-0.8% of viral particles in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, it is necessary to optimize the injection volume and viral titer to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral particle dosage in order to reduce the likelihood of Cre-independent expression.