Efficient introgression of allelic variants by embryo-mediated editing of the bovine genome.

摘要: The recent development of designer nucleases allows for the efficient and precise introduction genetic change into livestock genomes. Most studies so far have focused on random mutations in cultured cells use nuclear transfer to generate animals with edited genotypes. To circumvent intrinsic uncertainties inefficiencies we directed our efforts specific changes by homology-driven repair directly vitro produced embryos. Initially, injected zinc finger nuclease (ZFN)-encoding mRNA or DNA bovine zygotes verify cleavage activity at their target site within gene beta-lactoglobulin (LGB) detected ZFN-induced 30% 80% Next, precisely LGB sequence, co-injected ZFNs transcription activator-like effector (TALENs) oligonucleotides (ODNs). Analysis embryos showed targeted up 33% (ZFNs) 46% blastocysts. Deep sequence analysis selected revealed contributions allele can reach 100% which implies that genome editing zygote injections facilitate one-step generation non-mosaic pre-designed biallelic modifications.