CRISPR/Cas9高通量筛选技术与肿瘤治疗

Application of CRISPR/Cas9 High-throughput Screening Technology for Identifying New Target in Tumor Therapy

成簇规律间隔短回文重复(CRISPR),是细菌或古菌在与噬菌体长期生存进化获得的一种免疫系统。根据Cas蛋白(CRISPR-associated protein)的不同,CRISPR系统可分为3种。其中Ⅱ型CRISPR/Cas9已被改造成为一种有效的基因编辑工具,并运用于多种物种基因的改造。作为1种基因编辑的手段,CRISPR/Cas9技术通过诱导DNA双链断裂损伤,进一步干扰基因的表达。与传统的基因编辑技术相比,CRISPR/Cas9技术显示出效率高、成本低和易操作等特点。与此同时,二代测序技术的发展促进全基因组的解析。CRISPR技术结合高通量二代测序手段的使用,在肿瘤的治疗领域中已发挥出了独特的优势。本文就近年来CRISPR/Cas9高通量筛选技术的发展,及其在肿瘤治疗过程中的应用进行综述。

Clustered regularly interspaced short palindromic repeats（ CRISPR） is an acquired immunity system during the defensive processes of bacteria or archaea against bacteriophage. Basing on the difference about Cas proteins（ CRISPR-associated protein）,CRISPR system could be divided into three different types. Type Ⅱ CRISPR system-CRISPR/Cas9 has been modified to disturb gene function in various species. As a new gene editing tool,CRISPR/Cas9 technique could be employed to interrupt the expression of gene via triggering DNA double strand break（ DSB）. Compared with traditional gene modifying technologies,for example,RNA interference（ RNAi）,TALEN（ transcription activator-like effector nucleases）,and ZFN（ zinc finger nuclease）,CIRSPR/Cas9 system is possessed of some advantages such as high-efficiency,low-cost,and easy-operation. Additionally,rapid development of next-generation sequencing（ NGS） technology accelerates whole genome analysis. Therefore,CRISPR system combined with high-throughput NGS technology has displayed a unique advantage in tumor therapy. Hitherto there are three kinds of high-throughput tools based on CRISPR techniques,including CRISPR,CRISPR activation（ CRISPRa）,and CRISPR interference（ CRISPRi）. Here,we briefly summarized these three high-throughput screen strategies and discussed the related applications during different processes of cancer.