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FAQ

Frequently Asked Questions

 

Q: Can my gene of interest or expression cassette be packaged into AAV?

If your gene of interest is smaller than 4.9 kilo base pairs (kbp), we may package it into our rAAV products. We do not package gene sequences that may be unsafe to humans or the environment, such as those that are carcinogenic and those that encode for dangerous pathogens. If you have any further questions on this topic, you may contact us.

 

Q: What kind of AAV should I use for my tissue- or organ-specific studies?

Please see Introduction to AAV for pertinent background information and a useful chart detailing different serotypes of AAV. You may find one to fit your needs.

 

Q: What are the AAV 5.2 and AAV 8.2 products?

These AAV products are modifications of wild-type AAV5 and AAV8. In each product, the Phospholipase A2 domain encoded by the VP1 Cap gene has been replaced with the corresponding domain from AAV2. Functionally, the activity of this enzyme allows the virus to escape the cellular vesicle once it has been endocytosed into the target cell, therefore avoiding breakdown by lysozymes. AAV2 demonstrates some of the highest phospholipase activity, so we have integrated the AAV2 wild-type domain into our AAV5.2 and AAV8.2 products with no alteration in tropism, to increase infectivity.

 

Q: What do I send to Virovek before you can initiate my project?

Please see the section titled “Materials Required” on our How to Order page.

 

Q: I do not have ready-made DNA for my experiment. Can I still use your AAV Production Services?

Yes! We provide Gene Cloning Services for customers who have DNA sequence data, but no plasmids. This service is free for customers who want to proceed with our AAV Production.

 

Q: Is AAV safe to work with?

Yes, our rAAV is replication-defective, non-communicable, and ready for use in Biosafety Level 1 (BSL-1) settings. We express AAV rep and Cap genes in trans with our unique baculovirus expression system. Baculovirus is specific to insect cells and poses no threat to mammalian cell lines. Our technology allows us to make AAV products without the risk of residual adenovirus or herpes simplex virus contamination (these are both commonly used as helper viruses). Wild-type AAV already lacks many proteins necessary to reproduce on its own, making it a prime candidate for gene therapy research. For more information, please visit Safety and Storage, where you may view our Safety Data Sheet.

 

Q: How should I store the AAV that I receive?

AAV has been proven stable at 4° C and room temperature for use in immediate experiments. For long-term storage, please keep AAV at -80° C for best quality and longevity. Research suggests that storing AAV at -20° C is not sufficient for long-term preservation, and may reduce transduction efficiency of the virus. Be advised that although AAV is a very stable virus, multiple freeze-thaw cycles may reduce the titer of the product. For more information, please visit Safety and Storage and see our AAV Vector Storage and Handling Information.

 

Q: Can I reorder more of the same virus product with the same Lot Number?

Yes! Virovek keeps samples and remainders of custom-made AAV for customers up to 4 years after your initial order, to run assays and other troubleshooting tests. Please let us know what Lot Number and volume you would like to re-order, and we will provide you a quote. If you are interested in a titer adjustment, please let us know.

 

Q: How do you pack and ship your AAV products?

We pack our AAV tubes in a bubble mailer, surrounded by ice packs in a Styrofoam container and/or an outer cardboard box. The outside of the box will be clearly labeled with Virovek’s logo. Appropriate protocols on viral transduction will be included in the shipment. We ship through UPS WorldWide Express (WWEx). We encourage you to consider consolidating products from different quotes so we may ship them together in the same package.