CRISPR screen reveals modifiers of rAAV production including known rAAV infection genes playing an unexpected role in vector production.

ABSTRACT: Recombinant adeno-associated virus (rAAV) vectors are an effective and well-established tool in the growing gene therapy field, with five U.S. Food and Drug Administration-approved AAV-mediated gene therapies already on the market and numerous more in clinical trials. However, manufacturing rAAV vectors is an expensive, timely, and labor-intensive process that limits the commercial use of AAV-mediated gene therapies. To address this limitation, we screened producer cells for genes that could be targeted to increase rAAV yield. Specifically, we performed a CRISPR-based genome-wide knockout (KO) screen in human embryonic kidney (HEK) 293 cells using an antibody specific to intact AAV2 capsids coupled with flow cytometry to identify genes that modulate rAAV production. We discovered that the

SUBMITTER: O'Driscoll EE 

PROVIDER: S-EPMC11840188 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature