CRISPR/Cas9系统在昆虫中的应用

doi:10.16288/j.yczz.17-263

遗传 ›› 2018, Vol. 40 ›› Issue (4): 266-278.doi: 10.16288/j.yczz.17-263

CRISPR/Cas9系统在昆虫中的应用

童晓玲^1,²,方春燕^1,²,盖停停^1,²,石津^1,²,鲁成^1,²,代方银^1,²()

^1. 西南大学,家蚕基因组生物学国家重点实验室,重庆 400715
^2. 西南大学生物技术学院,农业部蚕桑生物学与遗传育种重点实验室,重庆 400715

收稿日期:2017-10-16 修回日期:2018-01-21 出版日期:2018-04-20 发布日期:2018-01-24
通讯作者: 代方银 E-mail:fydai@swu.edu.cn
作者简介:童晓玲,博士,副教授,硕士生导师,研究方向：昆虫发育遗传。E-mail: xltong@swu.edu.cn;方春燕,硕士研究生,专业方向：昆虫发育遗传。E-mail: 1039453631@qq.com;童晓玲和方春燕并列第一作者。|方春燕,硕士研究生,专业方向：昆虫发育遗传。E-mail: 1039453631@qq.com
基金资助:
国家高技术研究发展计划(863计划)项目(2013AA102507);国家自然科学基金项目(31472153);国家自然科学基金项目(31372379);现代农业产业技术体系专项资助(CARS-18)

Applications of the CRISPR/Cas9 system in insects

Tong Xiaoling^1,²,Fang Chunyan^1,²,Gai Tingting^1,²,Shi Jin^1,²,Lu Cheng^1,²,Dai Fangyin^1,²()

^1. State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400715, China
^2. Key Laboratory of Sericultural Biology and Genetic Breeding, Ministry of Agriculture, College of Biotechnology, Southwest University, Chongqing 400715, China

Received:2017-10-16 Revised:2018-01-21 Online:2018-04-20 Published:2018-01-24
Contact: Dai Fangyin E-mail:fydai@swu.edu.cn
Supported by:
Supported by the The National High Technology Research and Development Program of China (863 Program)(2013AA102507);the National Natural Science Foundation of China(31472153);the National Natural Science Foundation of China(31372379);the Technical System Special of Modern Agricultural industry of China(CARS-18)

摘要/Abstract

摘要：

CRISPR/Cas9(clustered regularly interspaced short palindromic repeat/CRISPR-associated nuclease 9)技术是一种RNA引导的基因组靶向编辑技术,能对基因组序列进行精确编辑,在探究基因功能、修复受损基因、沉默有害基因、改良品质性状等方面具有广阔的应用前景。近年来,随着对CRISPR/Cas9系统研究的不断深入和改造,该系统以其操作简易、省时、高效等优点在生物学研究的众多领域中得以推广和应用,特别是在果蝇(Bombyx mori)、家蚕(silkworm)、埃及伊蚊(Aedes aegypti)和蝴蝶(butterfly)等多种昆虫中。本文概述了CRISPR/Cas9的结构、作用原理及发展优化,总结了CRISPR/Cas9导入昆虫的策略和在昆虫中的应用,以及对CRISPR/Cas9系统产生脱靶问题的应对策略,以期对经济昆虫和有益昆虫的分子育种、害虫的生物技术防控等研究提供参考。

关键词: CRISPR/Cas9, 基因组编辑, 昆虫, 应用

Abstract:

The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated) system guides Cas9 to specific genomic locations by a short RNA search string. This technology enables the systematic interrogation of mammalian genome editing, repairing damaged genes, silencing harmful genes and improving quality traits. In recent years, with the introduction of the CRISPR/Cas9 system for easy, fast and efficient genetic modification, it has been possible to conduct meaningful functional studies in a broad array of insect species, such as Drosophila, Bombyx mori, Aedes aegypti and butterflies et al. In this review, we summarize the application of CRISPR/Cas9 in different insect species, discuss methods for its promotion, and consider its application for future insect studies.

Key words: CRISPR/Cas9, genome editing, insects, application

童晓玲,方春燕,盖停停,石津,鲁成,代方银. CRISPR/Cas9系统在昆虫中的应用[J]. 遗传, 2018, 40(4): 266-278.

Tong Xiaoling,Fang Chunyan,Gai Tingting,Shi Jin,Lu Cheng,Dai Fangyin. Applications of the CRISPR/Cas9 system in insects[J]. Hereditas(Beijing), 2018, 40(4): 266-278.

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[1]	郑晓飞,黄海燕,吴强. 染色质架构蛋白CTCF调控UGT1基因簇的表达[J]. 遗传, 2019, 41(6): 509-523.
[2]	王珏, 黄娟, 许蕊. 利用CRISPR/Cas9和piggyBac实现果蝇基因组无缝编辑[J]. 遗传, 2019, 41(5): 422-429.
[3]	吴凯,罗朝晖. 昆虫学案例在遗传学教学中的应用[J]. 遗传, 2019, 41(4): 349-358.
[4]	胡伟娟, 凌宏清, 傅向东. 植物表型组学研究平台建设及技术应用[J]. 遗传, 2019, 41(11): 1060-1066.
[5]	张楷, 刘蔚, 刘小凤, 陈瑶生, 刘小红, 何祖勇. 利用CRISPR/Cas9系统构建人HPRT1基因定点突变细胞株[J]. 遗传, 2019, 41(10): 939-949.
[6]	刘恒, 李东明, 朱兰玉, 赖乐锦, 闫婉云, 陆玉双, 韦伊, 黄月琪, 方媚, 苏元港, 杨芳, 舒伟. 利用CRISPR/Cas9敲除人源细胞系中LMNA基因的研究[J]. 遗传, 2019, 41(1): 66-75.
[7]	任云晓, 肖茹丹, 娄晓敏, 方向东. 基因编辑技术及其在基因治疗中的应用[J]. 遗传, 2019, 41(1): 18-27.
[8]	张桂珊, 杨勇, 张灵敏, 戴宪华. 机器学习方法在CRISPR/Cas9系统中的应用[J]. 遗传, 2018, 40(9): 704-723.
[9]	刘海龙, 谌阳, 高杨, 周玲, 韩晓松, 赵长志, 杨高娟, 陈毅龙, 杨慧, 谢胜松. 靶向miRNA前体不同类型sgRNA的丰度及特异性评估[J]. 遗传, 2018, 40(7): 561-571.
[10]	宋洁, 吴永波, 周跃恒, 柳波娟, 王楠, 郝转芳, 吴元奇. 作物组学数据库的比较和应用[J]. 遗传, 2018, 40(7): 534-545.
[11]	唐浚博, 曹浩伟, 许蕊, 张丹丹, 黄娟. 果蝇睾丸基因敲除突变体的构建及表型分析[J]. 遗传, 2018, 40(6): 478-487.
[12]	李慧卿, 陈超, 陈冉冉, 宋雪薇, 李佶娜, 朱延明, 丁晓东. 利用CRISPR/Cas9双基因敲除系统初步解析大豆GmSnRK1.1和GmSnRK1.2对ABA及碱胁迫的响应[J]. 遗传, 2018, 40(6): 496-507.
[13]	刘小民, 袁明龙. 昆虫天然免疫相关基因研究进展[J]. 遗传, 2018, 40(6): 451-466.
[14]	梁彩娇, 孟繁梅, 艾云灿. 基于CRISPR/Cas系统的噬菌体基因组编辑[J]. 遗传, 2018, 40(5): 378-389.
[15]	谢晶, 范辰, 张景龙, 张仕强. Ash2l-1/Ash2l-2在小鼠胚胎干细胞中的表达特异性及互补效应[J]. 遗传, 2018, 40(3): 237-249.