CAJ | 학술논문

基于 CRISPR/Cas9系统介导的第三代基因组编辑技术，已成功应用于动物、植物和微生物等诸多物种的基因组改造。如何提高 CRISPR/Cas9技术的基因组编辑效率和最大限度降低脱靶风险一直是本领域的研究热点，而使用高效且特异的sgRNA(Small guide RNA)是基因组改造成功的关键性因素之一。目前，已有多款针对 CRISPR/Cas9技术的 sgRNA 设计和/或脱靶效应评估软件，但不同的软件各有优缺点。本文重点对16款sgRNA 设计和脱靶效应评估在线和单机版软件的特点进行了阐述，通过制定38项评估指标对不同软件进行了比较分析，最后对11种用于检测基因组编辑效率和脱靶的实验方法，以及如何筛选高效且特异的sgRNA进行了归纳总结。
기우 CRISPR/Cas9계통개도적제삼대기인조편집기술，이성공응용우동물、식물화미생물등제다물충적기인조개조。여하제고 CRISPR/Cas9기술적기인조편집효솔화최대한도강저탈파풍험일직시본영역적연구열점，이사용고효차특이적sgRNA(Small guide RNA)시기인조개조성공적관건성인소지일。목전，이유다관침대 CRISPR/Cas9기술적 sgRNA 설계화/혹탈파효응평고연건，단불동적연건각유우결점。본문중점대16관sgRNA 설계화탈파효응평고재선화단궤판연건적특점진행료천술，통과제정38항평고지표대불동연건진행료비교분석，최후대11충용우검측기인조편집효솔화탈파적실험방법，이급여하사선고효차특이적sgRNA진행료귀납총결。
The third generation of CRISPR/Cas9-mediated genome editing technology has been successfully ap-plied to genome modification of various species including animals, plants and microorganisms. How to improve the efficiency of CRISPR/Cas9 genome editing and reduce its off-target effects has been extensively explored in this field. Using sgRNA (Small guide RNA) with high efficiency and specificity is one of the critical factors for successful ge-nome editing. Several software have been developed for sgRNA design and/or off-target evaluation, which have ad-vantages and disadvantages respectively. In this review, we summarize characters of 16 kinds online and standalone software for sgRNA design and/or off-target evaluation and conduct a comparative analysis of these different kinds of software through developing 38 evaluation indexes. We also summarize 11 experimental approaches for testing ge-nome editing efficiency and off-target effects as well as how to screen highly efficient and specific sgRNA.