Preventing packaging of translatable P5-associated DNA contaminants in recombinant AAV vector preps.

ABSTRACT: Recombinant adeno-associated virus (rAAV) vectors are increasingly being used for clinical gene transfer and have shown great potential for the treatment of several monogenic disorders. However, contaminant DNA from producer plasmids can be packaged into rAAV alongside the intended expression cassette-containing vector genome. The consequences of this are unknown. Our analysis of rAAV preps revealed abundant contaminant sequences upstream of the AAV replication (Rep) protein driving promoter, P5, on the Rep-Cap producer plasmid. Characterization of P5-associated contaminants after infection showed transfer, persistence, and transcriptional activity in AAV-transduced murine hepatocytes, in addition to in vitro evidence suggestive of integration. These contaminants can also be efficie

SUBMITTER: Brimble MA 

PROVIDER: S-EPMC8829444 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature