遗传 ›› 2020, Vol. 42 ›› Issue (3): 236-249.doi: 10.16288/j.yczz.19-297

• 综述 • 上一篇    下一篇

CRISPR-Cas基因编辑系统升级:聚焦Cas蛋白和PAM

唐连超, 谷峰()   

  1. 温州医科大学附属眼视光医院,眼视光学和视觉科学国家重点实验室,温州 325027
  • 收稿日期:2019-11-12 修回日期:2020-02-26 出版日期:2020-03-20 发布日期:2020-03-04
  • 通讯作者: 谷峰 E-mail:fgu@mail.eye.ac.cn
  • 作者简介:唐连超,硕士研究生,专业方向:基因编辑工具优化及应用。E-mail: 1664089980@qq.com
  • 基金资助:
    国家自然科学基金项目资助编号(81201181)

Next-generation CRISPR-Cas for genome editing: focusing on the Cas protein and PAM

Lianchao Tang, Feng Gu()   

  1. State Key Laboratory of Ophthalmology and Optometry, School of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou 325027, China
  • Received:2019-11-12 Revised:2020-02-26 Online:2020-03-20 Published:2020-03-04
  • Contact: Gu Feng E-mail:fgu@mail.eye.ac.cn
  • Supported by:
    Supported by the National Natural Science Foundation of China No(81201181)

摘要:

以CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR associated proteins)系统为代表的基因编辑技术的出现极大地促进了人类改造自然界物种的能力。在医疗、工业、农业等多个研究领域,基因编辑技术正在被广泛应用。Cas蛋白是CRISPR-Cas系统的功能蛋白,不同类型的Cas蛋白在其自身活性、识别位点、切割末端、RNA需求等方面具有不同的特性。PAM (protospacer adjacent motif)是靶位点附近的若干个碱基,对Cas蛋白识别靶序列至关重要,也是CRISPR-Cas系统发挥功效的关键特性之一。目前已有多种不同的PAM鉴定方法被报道。本文对Cas蛋白的寻找、Cas蛋白突变体筛选及PAM的确定方法(含PAM谱拓展)进行了综述,以期为新型基因编辑工具的发展和优化提供借鉴。

关键词: 基因编辑, CRISPR-Cas系统, Cas蛋白, PAM, 定向进化

Abstract:

The emergence of the gene editing technology, especial CRISPR-Cas (clustered regularly intersected short palindromic repeats and CRISPR associated proteins), has greatly promoted the ability of human beings to transform natural species. It has been widely harnessed for the engineering in the medical, industrial, agricultural and other fields. The key component of the CRISPR-Cas system, the Cas protein, possesses its specific features, including self-activity, recognition site, cutting end and guide RNA. PAM (protein assistant motif) is a number of nucleotides adjacent to the target site, which is very important for the Cas protein to recognize the target sequence and is also the key characteristic of CRISPR-Cas. There are several reported methods for identification of PAM. In this review, we summarize the searching for the Cas protein, the identification of Cas mutants with desired traits and the mapping of the PAM (including the extending of PAM spectrum), in order to provide a reference for the development and optimization of next-generation gene editing tools.

Key words: gene editing, CRISPR-Cas system, Cas protein, PAM, directed evolution