Single Cas9 nickase induced generation of NRAMP1 knockin cattle with reduced off-target effects

In this study, using chromatin immunoprecipitation and high-throughput sequencing (ChIP-seq), we identified the main binding sites of catalytically inactive Cas9 (dCas9) protein in bovine fetal fibroblast cells (BFFs) in an unbiased manner. Overall design: In the present study, chromatin immunoprecipitation was performed on catalytically inactive double mutant (D10A and H840A) Cas9 (dCas9)(Mali et al. 2013a; Yang et al. 2013) protein binding site detection in bovine fetal fibroblast cells (BFFs), followed by sequencing (ChIP-seq); we found that single Cas9 nickase (Cas9n)(Cong et al. 2013; Ran et al. 2013a)-mediated single-strand break (SSB) had the potential to restrict repair to the homology-directed repair (HDR) pathway.