摘要：Targeted mutagenesis by zinc-finger nucleases (ZFNs) can be used to generate knock-out mammalian cell lines with high efficiency. A number of different methods have been developed for the design and assembly of gene-specific ZFNs, making them easily accessible to researchers. In this study, we used ZFNs assembled through the CoDA (context-dependent assembly) platforms to generate mutant caprine fetal fibroblasts cells for the BLG gene. ZFN plasmid was introduced into the caprine fetal fibroblasts cell by electroporation. ZFN-induced cleavage of the target sequence was confirmed by the Surveyor nuclease assay analysis. Sequence analysis revealed that ZFN-induced mutations such as base insertion, deletion, or substitution were generated in the ZFN cleavage site of BLG. The simplicity and efficacy of CoDA will enable broad application of ZFN technology. This technique could be used with homologous arm, which may target foreign genes into the BLG locus at higher efficiency.
