遗传 ›› 2018, Vol. 40 ›› Issue (5): 378-389.doi: 10.16288/j.yczz.17-419

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基于CRISPR/Cas系统的噬菌体基因组编辑

梁彩娇,孟繁梅,艾云灿()   

  1. 中山大学生命科学学院,有害生物控制与资源利用国家重点实验室,广州 510275
  • 收稿日期:2017-12-25 修回日期:2018-02-06 出版日期:2018-05-20 发布日期:2018-04-04
  • 通讯作者: 艾云灿 E-mail:Lssayc@mail.sysu.edu.cn
  • 作者简介:梁彩娇,硕士研究生,研究方向:微生物学。E-mail: 515059960@qq.com
  • 基金资助:
    国家科技重大专项(2014ZX0801105B002);国家高技术研究发展计划(863计划)项目资助(2006AA09Z420)

CRISPR/Cas systems in genome engineering of bacteriophages

Caijiao Liang,Fanmei Meng,Yuncan Ai()   

  1. State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, China
  • Received:2017-12-25 Revised:2018-02-06 Online:2018-05-20 Published:2018-04-04
  • Contact: Ai Yuncan E-mail:Lssayc@mail.sysu.edu.cn
  • Supported by:
    Supported by the National Science and Technology Major Project of China(2014ZX0801105B002);and the National High Technology Research & Development Project(2006AA09Z420)

摘要:

对原核生物获得性免疫系统CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR- associated genes)的研究促进了新一代基因组编辑工具的产生和发展。噬菌体既是原核生物CRISPR阵列(CRISPR array)进化的原动力,又是CRISPR/Cas系统防御的对象。噬菌体功能基因组学研究的速率却落后于发现新噬菌体和测定基因组序列的速率。基于CRISPR/Cas系统的噬菌体基因组编辑,可为噬菌体功能基因组学研究提供新手段。本文评述了基于CRISPR/Cas系统编辑噬菌体基因组的几例开创性研究,并且比较了多种操作程序的异同点和优缺点。同时,进一步构建了联合使用CRISPR/Cas系统与噬菌体重组系统开展噬菌体基因组编辑的新方案,讨论了新方案的潜在局限性,并对如何选择不同方案给予了建议。

关键词: CRISPR/Cas系统, 噬菌体, 基因组编辑, 同源重组

Abstract:

Researches on CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated genes) systems, that are adaptive immunity systems encoded by prokaryotes, have promoted the development of new genome-editing tools. Bacteriophages are not only the driving elements for the evolution of prokaryotes’ CRISPR arrays, but also the targets of the CRISPR/Cas systems. Studies on functional genomics of bacteriophages have been lagging behind the discovery of new phage strains and the sequencing of their genomes. CRISPR/Cas systems-driven genome engineering of bacteriophages provides a novel approach for bacteriophage functional genomics. This review comments on a few profound cases of genome engineering of bacteriophages that employed the CRISPR/Cas systems, and compares multiple procedures illustrating common or distinct features as well as advantages and disadvantages underlying each procedure. We design new applications of the CRISPR/Cas systems coupled with bacteriophage recombination systems, discuss their potential constraints, and offer suggestions for each option.

Key words: CRISPR/Cas systems, bacteriophage, genome editing, homologous recombination