锌指蛋白核酸酶的作用原理及其应用钟强.pdfVIP

HEREDITAS (Beijing) 2011 2 , 33(2): 123―130 ISSN 0253-9772 综 述 DOI: 10.3724/SP.J.1005.2011.00123 锌指蛋白核酸酶的作用原理及其应用 钟强, 赵书红 , , 430070 锌指蛋白核酸酶(Zinc finger nucleases, ZFN) 因其能特异性识别并切割 DNA 序列以及可设计性, 被 用于基因定点突变和外源基因定点整合。目前, ZFN 技术以其准确的靶位点设计能力和诱发高效率基因打靶的 优势, 越来越受到基因改造研究者的重视, 已经成功应用于动植物细胞、胚胎的基因改造。随着鉴定靶 DNA 高亲和力的锌指蛋白(Zinc finger protein, ZFP)实验技术日渐成熟, 可以预见到不久的将来这项技术会在基因工 程和育种中得到广泛应用。文章介绍了锌指蛋白识别DNA 靶位点和ZFN 介导的基因打靶(Double strand break gene targeting, DSB-GT) 的原理, 同时还综述了目前ZFN 技术用于基因改造的研究进展。 锌指结构域; 锌指蛋白; 锌指蛋白核酸酶; 同源重组; 基因打靶 The mechanism and application of zinc finger nucleases ZHONG Qiang , ZHAO Shu-Hong Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education of China, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China Abstract: Due to specific recognization of DNA sequences and designability, zinc finger nucleases (ZFN) has been used in knock in and knock out genes. Because of high ratio of homologous recombination of DSB-GT induces by ZFN, ZFN technology has been considered as the most powerful tool for gene modification. It has been successfully used at cell or embryo levels in plants and animals. Under the rapid development of high affinity zinc finger protein (ZFP), this technique will be applied to genetic engineering and breeding extensively in the future. This review discussed the DNA recognization mechanism and double strand break gene targeting (DSB-GT) of zinc finger nucleases (ZFN). Some application examples of ZFN were summarized. Keywords: zinc finger domain; zinc finger protein; zinc finger nucleases; homologous recombination; gene targeting , nucleases, ZFN) DNA, , DNA