Highly efficient CRISPR/Cas9-mediated transgene knockin at the H11 locus in pigs

• DOI: 10.1038/srep14253
• Publication Date: 2015-09-18T09:04:36Z
• Authors: Jinxue Ruan; Hegang Li; Kui Xu; Tianwen Wu; Jingliang Wei; Rong Zhou; Zhiguo Liu; Yulian Mu; Shulin Yang; Hongsheng Ouyang; Ruby Yanru Chen-Tsai; Kui Li
• Keywords: Gene targeting

Abstract Transgenic pigs play an important role in producing higher quality food agriculture and improving human health when used as animal models for various diseases biomedicine. Production of transgenic pigs, however, is a lengthy inefficient process that hinders research using pig models. Recent applications the CRISPR/Cas9 system generating site-specific gene knockout/knockin models, including knockout model, have significantly accelerated model field. However, knockin containing transgene insertion can be passed on to its offspring remains lacking. Here, we describe first time generation combination somatic cell nuclear transfer. We also report new genomic “safe harbor” locus, named pH11 , which enables stable robust expression. Our results indicate our allows highly efficient at locus up 54% drug selection 6% without selection. successfully inserted fragment larger than 9 kb system. data confirm into expressed cells, embryos animals.