微型CRISPR核酸酶Cas12f研究进展及在基因编辑领域中的应用  

Research progress of miniature CRISPR nuclease cas12f and its application in the field of gene editing

作　　者：刘亭君 陈祥凤 王俊利[2] 林敏[3] 董明右 LIU Ting-jun;CHEN Xiang-feng;WANG Jun-li;LIN Min;DONG Ming-you(School of Laboratory Medicine,Youjiang Medical University for Nationalities,Baise 533000,China;Modern Industrial College of Biomedicine and Great Health,Youjiang Medical University for Nationalities,Baise 533000,China;School of Life Sciences and Food Engineering,Hanshan Normal University,Chaozhou 521041,China)

机构地区：[1]右江民族医学院医学检验学院,百色533000 [2]右江民族医学院现代产业学院,百色533000 [3]韩山师范学院生命科学与食品工程学院,潮州521041

出　　处：《中国人兽共患病学报》2023年第2期172-179,共8页Chinese Journal of Zoonoses

基　　金：广西自然科学基金面上项目(No.2020JJA140656);广东省教育厅科研项目(No.2019-GDXK-0031);广西高校中青年教师科研基础能力提升项目(No.2022KY0532)联合资助。

摘　　要：成簇的规律间隔的短回文重复序列及其相关蛋白(CRISPR-Cas)系统是一种存在于大多数细菌和古菌中的“获得性免疫系统”,已被广泛应用于生物、医学等多个研究领域。传统的Cas核酸酶(Cas9、Cas12和Cas13)因为分子尺寸较大,将基因编辑工具精确送到真核细胞内受到阻碍。研究发现,Cas12f是目前已知的最小RNA引导核酸酶,有着与众不同的属性:除分子比较小外;对单链DNA切割不需要靶标前间区序列邻近基序(Protospacer adjacent motif,PAM);非常长的(153~169个核苷酸)反式激活crRNA(tracrRNA);结合单拷贝向导RNA(gRNA)后的二聚化;不对称的双链切割模式;针对单链DNA的附带切割活性;对识别序列中间的碱基要求严格等。基于目前研究,Cas12f具体作用机制尚不明确,本文对近年来CRISPR-Cas12f起源、分类、作用机制及其在基因编辑领域中应用的研究进行综述,以期开发微型Cas12f精准基因编辑和治疗工具,同时为相关研究领域的科研工作提供参考。The clustered regularly interspaced short palindromic repeats and their associated proteins(CRISPR-Cas)system is an"acquired immune system"existing in most bacteria and archaea,and it has been widely used in many research fields such as biology and medicine.Due to the large molecular size of the traditional Cas nucleases(Cas9,Cas12 and Cas13),it is very difficult to deliver gene editing tools precisely into eukaryotic cells.It was found that Cas12f is the smallest RNA-guided nuclease known,with distinctive properties:relatively small molecular weight;no need for a target pre-intermediate sequence adjacent motif(PAM)for single-stranded DNA cleavage;very long(153-169 nucleotides)trans-activated crRNA(tracrRNA);dimerization upon binding of single-copy guide RNA(gRNA);asymmetric double-stranded cleavage pattern;incidental cleavage activity against single-stranded DNA;stringent requirements for bases in the middle of the recognition sequence,etc.Based on current studies,the specific mechanism of action of Cas12f is still unclear.This paper reviews the recent research literature on the origin,classification and mechanism of action of CRISPR-Cas12f and its application in the field of gene editing,with a view to developing miniature Cas12f precision gene editing and therapeutic tools,as well as providing a reference for scientific research in related research fields.

关 键 词：CRISPR Cas12f 单链DNA 基因编辑 

分 类 号：R446[医药卫生—诊断学]