Short DNA Hairpins Compromise Recombinant Adeno-Associated Virus Genome Homogeneity.

ABSTRACT: Short hairpin (sh)RNAs delivered by recombinant adeno-associated viruses (rAAVs) are valuable tools to study gene function in vivo and a promising gene therapy platform. Our data show that incorporation of shRNA transgenes into rAAV constructs reduces vector yield and produces a population of truncated and defective genomes. We demonstrate that sequences with hairpins or hairpin-like structures drive the generation of truncated AAV genomes through a polymerase redirection mechanism during viral genome replication. Our findings reveal the importance of genomic secondary structure when optimizing viral vector designs. We also discovered that shDNAs could be adapted to act as surrogate mutant inverted terminal repeats (mTRs), sequences that were previously thought to be required for functiona

SUBMITTER: Xie J 

PROVIDER: S-EPMC5474962 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature