基因组编辑技术的原理及应用  

Principle and application of the genome editing technology

作　　者：贺飞燕[1,2] 闫建俊[2,3] 冯瑞云[2] 张爱萍[4] 张维锋[2] 白云凤[2,3] 

机构地区：[1]山西大学生物工程学院,太原030006 [2]山西省农业科学院作物科学研究所,太原030006 [3]农业部黄土高原作物基因资源与种质创制重点实验室,太原030031 [4]新疆生产建设兵团第6师农业科学研究所,五家渠831300

出　　处：《应用与环境生物学报》2016年第2期350-356,共7页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(30971838);山西省科技攻关项目(201303110015-1);山西省回国留学人员基金项目(2013-146)

摘　　要：阐明基因功能和改良生物现状是生物学重点研究内容之一.近年来利用的基因打靶和转基因技术存在效率低、耗时长、易引起人们的安全性疑虑等问题.新近发现的以多种新型高效的DNA靶向内切酶为基础而建立的基因组编辑技术,主要包括ZFN、TALEN和CRISPR/Cas三种.本文首先综述这3种技术的原理,即均基于"DNA断裂/DNA损伤修复"来实现编辑功能,可以在不同物种中对目标基因进行定点敲除、单核苷酸或多核苷酸片段的置换、添加等基因组靶向修饰,具有简单、快速、高效、准确等优点.然后比较3种技术在构成、靶点识别模式、编辑特点、技术难度以及脱靶效应等方面的异同;归纳该技术在作物遗传育种、家畜改良以及基因治疗等方面的应用现状和前景,并指出该技术尚存在的脱靶、构建等问题以及解决的途径,重点突出ZFN、TALEN和CRISPR/Cas在模式植物以及粮食作物中的应用.最后提出建立基于病毒载体的基因组无痕编辑技术平台,将使基因组编辑用于植物遗传改良更为简约和安全.Elucidating gene function and improving biological status is one of the key research contents of biology. The use of gene targeting and transgenic technology is accompanied with problems including low efficiency, long time, safety doubts and others. New genome editing techniques based on novel and efficient DNA targets include ZFN, TALEN and CRISPR/Cas. In this paper, we first review the three technology principles, which all based on DNA/DNA damage repair to achieve editing function, can perform target gene deletion, single nucleotide or polynucleotide fragments replacement, and target gene modification in different species of the target gene. The new technology is simple, rapid, efficient and accurate. Then we compare the three techniques in terms of composition, target recognition mode, editing features, technical difficulty and offtarget effect. We also summarize the status and prospects of the application of this technology in crop genetics and breeding, livestock improvement and gene therapy. The review points out the shortcomings of the technology including off-target effect and construction problems, and provides with possible solutions. The application of ZFN, TALEN and CRISPR/Cas in the model plant and grain crops is emphasized. At the end of this paper, we also introduce the present research situation and our research ideology concerning the technology, proposing the establishment of the technology platform of genome editing based on viral vector, which will make simpler and safer application of genome editing for plant genetic improvement.

关 键 词：基因组编辑 ZFN TALEN CRISPR/Cas 原理 分子育种 

分 类 号：Q78[生物学—分子生物学]