Efficient modification of CCR5 in primary human hematopoietic cells using a megaTAL nuclease and AAV donor template.

ABSTRACT: Genetic mutations or engineered nucleases that disrupt the HIV co-receptor CCR5 block HIV infection of CD4(+) T cells. These findings have motivated the engineering of CCR5-specific nucleases for application as HIV therapies. The efficacy of this approach relies on efficient biallelic disruption of CCR5, and the ability to efficiently target sequences that confer HIV resistance to the CCR5 locus has the potential to further improve clinical outcomes. We used RNA-based nuclease expression paired with adeno-associated virus (AAV)-mediated delivery of a CCR5-targeting donor template to achieve highly efficient targeted recombination in primary human T cells. This method consistently achieved 8 to 60% rates of homology-directed recombination into the CCR5 locus in T cells, with over 80% of cel

SUBMITTER: Sather BD 

PROVIDER: S-EPMC4790757 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature