CRISPR_Cas9系统的分子制及其在人类疾病基因治疗中的应用要素.pdf

Hereditas (Beijing) 2015 10 , 37(10): 974 982 CRISPR/Cas9 CRISPR/Cas CRISPR/Cas 215123 CRISPR/ Cas9 CRISPR/Cas9 CRISPR/Cas9 The molecular mechanism of CRISPR/Cas9 system and its application in gene therapy of human diseases Liang Qu, Huashan Li, Yunhan Jiang, Chunsheng Dong Institutes of Biology and Medical Sciences (IBMS ), School of Medicine, Soochow University, Suzhou 215123, China Abstract: CRISPR/Cas system is an adaptive immune system that confers resistance to exogenous virus or plasmid in bacteria and archaea. In recent years, the booming CRISPR/Cas9 genome editing technology modified from type CRISPR/Cas adaptive immune system has been widely applied to various research fields of life science and led to revolutionary changes. In this review, we summarize the origin and development of CRISPR/Cas9 genome editing technology as well as its applications in life science research. We focus on the latest application of this system in gene therapy of human diseases and the associated side/off-target effects, which may provide references for researchers in related areas. Keywords: CRISPR/Cas9 system; adaptive immune system; genome editing 20 80 (Homologous recombination, HR) [1,2] : 2015-03-16; : 2015-07-23 : ( K113415011) SYS201452) : ( ) E-mail: quliangsuda@163.com : E-mail chunshengdong@ DOI: 10.16288/j.yczz.15-109 : 2015-8-4 9:37:07 URL: /kcms/detail/11.1913.R0937.004.html ( 201310285046Z) ( 10 : CRISPR/Cas9 975 21 (Zinc finger nuclease, ZFN) [3,4] (Transcription activator-like effector nucleases, TALEN) [5~7] DNA (Homology directed repair, HDR) DNA (Non-homologous end joining, NHEJ) DNA 2013 Science Nature Biotechnology Cell Fok —Clustered regularly interspaced short palindromic repeats /associated nuclease 9 (CRISPR/Cas9) CRISPR/Cas CRISPR/Cas9 [8~11] CRISPR/Cas9 (1) 20 bp sgRNA DNA ZFN TALEN DNA TALEN 7~9 1000 bp DNA (2) Cas9 RNA (Single-guide RNA sgRNA) DNA (3) CRISPR/Cas9 CRISPR/Cas9 Cas Cas10 AB A mRNA [14] B DNA [15] Type —Cas9 Cas9 crRNA [16,17