Adeno-Associated Virus Serotype 3 (AAV3) shares 82% sequence homology with AAV2, and like AAV2, requires Heparan Sulfate Proteoglycan (HSPG) for cellular 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 domain and the intracellular kinase domain of hHGFR are required for AAV3-mediated transgene expression.
These AAV3 particles constitutively express ZsGreen under a CMV promoter. ZsGreen is a human codon-optimized variant of the green fluorescent protein isolated from reef coral (Zoanthus sp). It has been engineered for higher expression in mammalian cells and is up to four times brighter than enhanced GFP (eGFP). ZsGreen expression and AAV3 transduction efficiency can easily be verified and optimized by fluorescence microscopy or flow cytometry. ZsGreen has an excitation wavelength of 493 nm and an emission wavelength of 505 nm.
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). The purity will vary with each lot; the exact value is provided with each shipment.
Formulation
AAV3 was produced in HEK293-AAV cells and is supplied in PBS-MK (PBS Magnesium-Potassium) buffer with 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 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.
Troubleshooting Guide For all further questions, please email [email protected].