AAV Comparison and Selection Using qPCR Methods

AAV vectors are powerful tools for gene delivery and gene therapy. Hundreds of identified natural AAV serotypes and increasing number of engineered variants allow us to find better capsids for each cell type and tissue, which is vital for effective gene therapies. However, there has been no good method to achieve that. Currently, the common way to compare AAV variants is to test them side-by-side individually, whereas it is difficult to generate, control and handle hundreds of AAV vectors in every lab or company while limited candidates decrease the chance to find better AAV variants.

To solve this problem, AAVnerGene provides AAV capsid selection services, which will help customers fastly and semi-high-throughputly compare the AAV variants both in vivo and in vitro.​​
 

Design of qPCR Based Capsid Selection

In order to efficiently compare the transduction efficiency of multiple AAV variants, each capsid variant packages a DNA barcoded  genome carrying a reporter gene(such as EGFP). The unique barcode is located between coding sequence and Poly-A signal. Then, the capsid/barcoded-transgene pairs were transfected into HEK 293 cells to produce AAV vectors. 

For less AAV variants(<24), the capsid/barcoded-transgene pairs are transfected and purified independently. After titration, equal amount of AAV vectors are pooled together and generate the final AAV vectors pool.