Novel lentiviral vectors with mutated reverse transcriptase for mRNA delivery of TALE nucleases.

ABSTRACT: TAL-effector nucleases (TALENs) are attractive tools for sequence-specific genome modifications, but their delivery still remains problematic. It is well known that the presence of multiple sequence repeats in TALEN genes hampers the use of lentiviral vectors. We report that lentiviral vectors readily package full-length vector mRNAs encoding TALENs, but recombination during reverse transcription prevents successful delivery. We reasoned that preventing reverse transcription of lentiviral-vector RNA would allow transfer of TALENs as mRNA. We demonstrate that lentiviral particles containing genetically inactivated reverse transcriptase (RT) mediated efficient transduction of cultured cells and supported transient transgene expression. For proof-of-principle, we transferred CCR5- and TCR-spe

SUBMITTER: Mock U 

PROVIDER: S-EPMC4166709 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature