Optimization of AAV6 transduction enhances site-specific genome editing of primary human lymphocytes.

ABSTRACT: Adeno-associated virus serotype 6 (AAV6) is a valuable reagent for genome editing of hematopoietic cells due to its ability to serve as a homology donor template. However, a comprehensive study of AAV6 transduction of hematopoietic cells in culture, with the goal of maximizing ex vivo genome editing, has not been reported. Here, we evaluated how the presence of serum, culture volume, transduction time, and electroporation parameters could influence AAV6 transduction. Based on these results, we identified an optimized protocol for genome editing of human lymphocytes based on a short, highly concentrated AAV6 transduction in the absence of serum, followed by electroporation with a targeted nuclease. In human CD4⁺ T cells and B cells, this protocol improved editing rates up

SUBMITTER: Rogers GL 

PROVIDER: S-EPMC8517001 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature