基因组编辑技术与模式动物

Genome editing technologies and model animal

随着高通量测序技术的发展,后基因组时代的研究重点已转移至如何阐明基因功能。以胚胎干细胞(ES)和同源重组为基础的基因打靶技术在生命科学及医学研究中做出了重要贡献,但存在打靶效率低、耗时长、有物种限制等缺点。近年来,锌指核酸酶(ZFNs)、类转录激活因子核酸酶(TALENs)及CRISPR/Cas9等新的基因组编辑技术的兴起极大地推进了基因功能研究的进展。ZFNs与TALENs包含两个结构域,识别并结合核苷酸的DNA结合域和Fok I核酸内切酶,两者的区别在其DNA识别结构域:ZFNs识别的基本单位为3个连续碱基对,TALENs则识别单个碱基。与ZFNs及TALENs不同,CRISPR/Cas9以碱基互补配对机制识别并结合DNA,该技术设计简便,在各物种中得到广泛应用。自ES细胞打靶技术至今,研究者采用基因打靶技术已获得许多疾病相关模型,这些模型在基因功能、人类疾病治疗及基因治疗等方面发挥着越来越重要的作用。将对各类基因组编辑技术的原理及应用情况进行总结,同时介绍模式动物在生命科学及医学研究中的应用情况。

With the development of high-throughput sequencing in the post genome era, the researchers concentrate their efforts on elucidating the relationships between the gene and the corresponding physiological/pathological function. The gene targeting technology based on the embryonic stem cells （ES） and homologous recombination technique has made a great contribution to the life science and medical research, meanwhile has the disadvantage of low efficiency, long time consumption and species limitation. Recently, the grown-up of new genome editing technology including ZFN, TALEN andCRISPR/Cas9 accelerates the progress of gene function study. ZFNs and TALENs both contain two domains： one executes the DNA recognition and binding function, the other is the Fok I endonuclease domain. The difference between their DNA binding domains is that ZFNs recognizes 3 successive bases as the basic unit while TALENs identifies the single base. Unlike ZFNs and TALENs, CRISPR/Cas9 recognizes and binds the target DNA based on the principle of complementary base pairing, which promotes the wide application of Cas9 in various species. Since the foundation of ES cell targeting technology, researchers have made many disease-related models using this technology, and these models play a more and more important role in human disease treatment, gene function study and gene therapy. Here, we summarized the principle and application of these genome editing technologies and introduced the application of animal model in the life science and medical research.