郭全娟,韩秋菊,张建.CRISPR/Cas9技术的脱靶效应及优化策略*[J].生物化学与生物物理进展(待发表)
CRISPR/Cas9技术的脱靶效应及优化策略*
Review: Off-target Effect of CRISPR / Cas9 and Optimization*
投稿时间:2018-01-09  修订日期:2018-05-23
中文关键词:  基因编辑,CRISPR/Cas9,脱靶效应,向导RNA
英文关键词:gene  editing, CRISPR/Cas9, off-target  effects, single-guideRNA
基金项目:国家自然科学基金(81373222, 81172789),山东省重点研发计划(2017GSF18159),山东省自然基金博士基金(ZR2017BH029)资助项目
作者单位E-mail
郭全娟 山东大学药学院免疫药物学研究所 gqj2018@163.com 
韩秋菊 山东大学药学院免疫药物学研究所 hanqiuju@sdu.edu.cn 
张建 山东大学药学院免疫药物学研究所 zhangj65@sdu.edu.cn 
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中文摘要:
      近年CRISPR/Cas9系统已经迅速成为医学领域最具潜力的基因编辑工具,它是继锌指核酸内切酶(Zinc-finger nucleases, ZFNs)和转录激活子样效应因子核酸酶(Transcription activator-like effectors nucleases, TALENs)之后的第三代人工改造的核酸内切酶,被喻为“基因魔剪”.该基因编辑系统最大优势在于只需要设计一个靶向目标序列的sgRNA(single-guide RNA)就能引导Cas9核酸酶结合到特定的基因序列,并指导 Cas9核酸酶切割相应的结合位点,进而完成对靶基因的编辑.由于其设计简单、实施快捷、成本低廉,突变效率高等优点,这一系统在基因组功能鉴定、抗病毒、抗肿瘤、免疫治疗等众多领域已广泛研究.虽然CRISPR/Cas9系统具有如此多的优点,但是其可能存在的脱靶风险影响了该基因编辑技术的深入研究,增加了用于疾病治疗的风险,阻碍了临床的进一步应用.如何保证CRISPR/Cas9基因编辑技术高编辑效率,同时也能降低脱靶效应,这将是未来科学研究者深入研究CRISPR/Cas9技术并开拓其临床应用亟待解决的问题.为此我们在文章中对CRISPR/Cas9系统组成、作用机制及应用进展进行简要综述,着重介绍目前这一领域广受关注的脱靶效应,并归纳优化策略,以期为更多的研究者提供参考.
英文摘要:
      In recent years, CRISPR/Cas9 system has rapidly become the most revolutionary gene editing tool in the field of medicine. It is the third-generation artificial endonuclease after Zinc-finger nucleases (ZFNs) and Transcription activator-like effectors nucleases (TALENs), and is known as "gene scissors". CRISPR/Cas9 system relies on a synthetic single guided RNA (sg RNA) to direct Cas9 nuclease to cleave the target DNA sequence, so that it can complete genomic site editing. Based on the simplicity of design, rapid implementation, low cost and high efficiency, CRISPR/Cas9 system has been extensively studied in the fields of gene functional identification, antiviral therapy, anti-tumor therapy and immunotherapy. Although CRISPR/Cas9 system displays several obvious advantages, the risk of off-target would increase the therapeutic risks, hinder research and clinical application, which should not be ignored. How to reduce the off-target effects and increase mutation efficiency simultaneously as we use CRISPR/Cas9 gene editing technology? This will be an urgent problem for scientists to explore CRISPR/Cas9 technology and expand its clinical application in the future. Here, we briefly introduce the composition, mechanism and application of CRISPR/Cas9 system, and give a comprehensive introduction of the off-target effect which is widely concerned in this field at present, and summarize the optimization strategies to provide references for researchers.
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