Zinc finger protein-dependent and -independent contributions to the in vivo off-target activity of zinc finger nucleases.

ABSTRACT: Zinc finger nucleases (ZFNs) facilitate tailor-made genomic modifications in vivo through the creation of targeted double-stranded breaks. They have been employed to modify the genomes of plants and animals, and cell-based therapies utilizing ZFNs are undergoing clinical trials. However, many ZFNs display dose-dependent toxicity presumably due to the generation of undesired double-stranded breaks at off-target sites. To evaluate the parameters influencing the functional specificity of ZFNs, we compared the in vivo activity of ZFN variants targeting the zebrafish kdrl locus, which display both high on-target activity and dose-dependent toxicity. We evaluated their functional specificity by assessing lesion frequency at 141 potential off-target sites using Illumina sequencing. Only a minorit

SUBMITTER: Gupta A 

PROVIDER: S-EPMC3017618 | biostudies-literature | 2011 Jan

REPOSITORIES: biostudies-literature