Adeno-Associated Virus Serotype 3 (AAV3) shares 82% sequence homology with AAV2, and like AAV2, requires the Heparan Sulfate Proteoglycan (HSPG) receptor for cell attachment. AAV3 vectors transduce human liver cancer cells extremely efficiently because AAV3 utilizes the human Hepatocyte Growth Factor Receptor (hHGFR) as a co-receptor for viral entry, which is highly expressed in these cells. Both the extracellular and intracellular kinase domains of hHGFR are required for AAV3-mediated transgene expression.
These AAV particles constitutively express the firefly (Photinus pyralis) luciferase and mCherry genes connected via a T2A linker, under the control of a CMV promoter. The T2A self-cleaving peptide (derived from Thosea asigna virus 2A) leads to the efficient cleavage of the transcript, and expression of luciferase and mCherry as two separate proteins.
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Product Data Gallery
Purified AAV3 Luciferase-mCherry particles.
Transduction of HEK293 cells using AAV3 Luciferase-mCherry particles.
Luciferase activity of HEK293 cells transduced by AAV3 Luciferase-mCherry particles.
Two vials (50 µl x 2) of AAV at a titer ≥1 x 1012 vector genomes/ml. The titer is determined by qPCR and will vary with each lot; the exact value will be provided with each shipment.
Purification
The purity of the AAV particles was confirmed to be greater than 90% by staining with One-Step Lumitein™ UV Protein Gel Stain (Biotium #21005-1L). Purity will vary with each lot; the exact value will be provided with each shipment.
Formulation
AAV3 was produced in HEK293-AAV cells and is supplied in PBS-MK (PBS Magnesium-Potassium) buffer containing 0.01% Pluronic F68.
Storage/Stability
AAV is shipped with dry ice. For long-term storage, it is recommended to store AAV at -80°C for up to 12 months from date of receipt. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle
Applications
Use as a positive control for transduction
Optimize transduction assays and track protein expression over time
Shipping Temperature
-80°C
Notes
Biosafety Recombinant AAV is inherently replication-deficient and not known to cause any human diseases. Additionally, following transduction, AAV vectors exist episomally and do not integrate into or disrupt the host cell’s genome. AAV requires the use of a Biosafety Level 1 facility. BPS Bioscience recommends following all local, federal, state, and institutional regulations and using all appropriate safety precautions.