基于体细胞克隆的猪基因打靶技术研究进展

Gene-targeted pigs based on somatic cell cloning approaches

基因打靶猪在农业和生物医药领域均有广泛用途。由于猪的能参与生殖系嵌合体的多能性干细胞尚未建立成功,基因打靶猪的培育主要是通过体细胞克隆技术来实现。最初,人们在体细胞上通过传统的同源重组技术成功地建立了基因敲除克隆猪,但体细胞在体外的增殖能力有限,传统同源重组在体细胞的打靶效率极低。虽经十多年的发展,但在世界范围内获得的基因打靶猪屈指可数。最近,三种工程核酸酶介导的基因编辑技术(ZFN、TALEN和CRISPR/Cas9)的出现,使体细胞的基因打靶效率大大提高。多个实验室将其用于猪,实现了高效基因打靶,并在很短的一段时间里,获得了一系列基因打靶猪。就传统同源重组技术以及近几年发展起来的新兴基因编辑技术在基因修饰猪的应用研究进展进行了综述。

Gene-targeted pigs have wide applications in agriculture and biomedicine. Due to unavailability of germ line competent pluripotent cells, the production of gene-targeted pigs is mainly through somatic cell cloning. Initially, homologous recombination technology was used to target the genome in pigs. However, the efficiency of gene targeting in somatic cells is extremely low because of＇ their low proliferative capacity in vitro. As a result, only a few gene-targeted pigs have been reported in last decade. With the three newly emerging gene editing technologies （ZFN, TALEN and CRISPR/CAS9） coming into use, the efficiency of gene targeting in somatic cells hassignificantly been improved. These three technologies have been successfully applied to pigs with a very high efficiency and many gene-targeted pigs have been generated. In this review, we discussed the progress of genetargeted pigs with traditional homologous recombination and newly emerging gene editing technologies.