基于CRISPR/Cas9系统在全基因组范围内筛选功能基因及调控元件研究进展

doi:10.16288/j.yczz.19-390

遗传 ›› 2020, Vol. 42 ›› Issue (5): 435-443.doi: 10.16288/j.yczz.19-390

基于CRISPR/Cas9系统在全基因组范围内筛选功能基因及调控元件研究进展

刘思远^1,², 易国强², 唐中林^2,³(), 陈斌¹()

^1. 湖南农业大学动物科学技术学院,长沙 410128
^2. 中国农业科学院农业基因组研究所,岭南现代农业科学与技术广东省实验室深圳分中心,农业部农业基因数据分析重点实验室,深圳 518120
^3. 中国农业科学院北京畜牧兽医研究所,北京 100193

收稿日期:2020-01-02 修回日期:2020-04-08 出版日期:2020-05-20 发布日期:2020-04-26
通讯作者: 唐中林,陈斌 E-mail:tangzhonglin@caas.cn;chenbin7586@126.com
作者简介:刘思远,在读博士研究生,专业方向：动物遗传育种与繁殖。E-mail: 515970802@qq.com|易国强,研究员,研究方向：功能基因组和表观遗传学。E-mail:yiguoqiang@caas.cn. 刘思远和易国强为并列第一作者。
基金资助:
湖南省生猪产业技术体系岗位专家项目编号(2019-2021);深圳市技术攻关项目编号(JSGG20180507182028625);转基因生物新品种培育重大专项编号(2016ZX08006002-005);广东省重点领域研发计划(现代种业)项目编号资助(2018B020203002)

Progress on genome-wide CRISPR/Cas9 screening for functional genes and regulatory elements

Siyuan Liu^1,², Guoqiang Yi², Zhonglin Tang^2,³(), Bin Chen¹()

^1. College of Animal Science & Technology, Hunan Agricultural University, Changsha 410128, China;
^2. Genome Analysis Laboratory of the Ministry of Agriculture, Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, China
^3. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2020-01-02 Revised:2020-04-08 Online:2020-05-20 Published:2020-04-26
Contact: Tang Zhonglin,Chen Bin E-mail:tangzhonglin@caas.cn;chenbin7586@126.com
Supported by:
Supported by Position Expert of Hunan Province Pig Industry Technology System No(2019-2021);Shenzhen Key Technology Projects No(JSGG20180507182028625);National Science and Technology Major Project of China No(2016ZX08006002-005);the Key R&D Programmes of Guangdong Province No(2018B020203002)

摘要/Abstract

摘要：

CRISPR/Cas9系统是一种近年来被广泛应用于基因组编辑的强大工具。通过将CRISPR/Cas9系统中的Cas9蛋白突变后,使其失去剪切活性而成为dCas9 (nuclease-dead Cas9),再结合基因功能丧失(loss-of-function, LOF)、基因功能激活(gain-of-function, GOF)以及非编码功能基因鉴定技术即可实现全基因组高通量的功能基因及调控元件靶向鉴定和筛选。目前,该技术已被广泛应用于疾病免疫机理、药物靶点筛选和动物遗传育种等研究,为生命医学和基础科学带来了全新高效的技术方法和研究思路。本文综述了基于CRISPR/Cas9技术在全基因组中高通量筛选功能基因及调控元件的方法及研究进展,重点阐述了CRISPR/Cas9系统在动物细胞中筛选功能性基因的方法,以期为基因编辑及相关研究领域提供参考。

关键词: CRISPR/Cas9, 全基因组筛选, 功能基因, 调控元件

Abstract:

The CRISPR/Cas9 system is a powerful tool which has been extensively used for genome editing in the past few years. Nuclease-dead Cas9 (CRISPR/dCas9), a Cas9 protein mutant without splicing ability, along with loss-of- function (LOF), gain-of-function (GOF), or non-coding genes scanning approaches can reveal genome-scale functional determinants. CRISPR/Cas9 has been widely adopted to decipher disease mechanisms and pinpoint drug targets in the life science field, and also provide novel insights into animal genetics and breeding. In this review, we summarize the research progress in high-throughput CRISPR/Cas9 screening for revealing the functional genes and regulatory elements in the whole genome. We also highlight the applications of CRISPR/Cas9 system in the animal cells, providing a reference for gene editing and other related research in related fields.

Key words: CRISPR/Cas9, genome-wide screening, functional gene, regulatory elements

刘思远, 易国强, 唐中林, 陈斌. 基于CRISPR/Cas9系统在全基因组范围内筛选功能基因及调控元件研究进展[J]. 遗传, 2020, 42(5): 435-443.

Siyuan Liu, Guoqiang Yi, Zhonglin Tang, Bin Chen. Progress on genome-wide CRISPR/Cas9 screening for functional genes and regulatory elements[J]. Hereditas(Beijing), 2020, 42(5): 435-443.

图/表 1

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基于CRISPR/Cas9系统在全基因组范围内筛选功能基因及调控元件研究进展

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