从随机突变到精确编辑：果蝇基因组编辑技术的发展及演化

doi:10.16288/j.yczz.15-337

遗传 ›› 2016, Vol. 38 ›› Issue (1): 17-27.doi: 10.16288/j.yczz.15-337

从随机突变到精确编辑：果蝇基因组编辑技术的发展及演化

苏方¹, 黄宗靓¹, 郭雅文², 焦仁杰^{1, 2}, 李孜¹, 陈建明^{1, 3}, 刘继勇¹

1. 广州医科大学广州霍夫曼免疫研究所, 广州 511436;
2. 中国科学院生物物理研究所脑与认知科学国家重点实验室,北京 100101;
3. 国家海洋局第三海洋研究所,厦门 361005

收稿日期:2015-07-27 修回日期:2015-10-19 出版日期:2016-01-20 发布日期:2016-01-20
通讯作者: 刘继勇,博士,副教授,研究方向：果蝇天然免疫。E-mail: mangriver@hotmail.com;陈建明,博士,教授,研究方向：血细胞进化与适应性。E-mail: chenjm@xmu.edu.cn;李孜,博士,教授,研究方向：果蝇天然免疫。E-mail: lizi1002@gzhmu.edu.cn E-mail:fx-su@126.com
作者简介:苏方,硕士,专业方向：果蝇天然免疫。
基金资助:
国家自然科学基金青年基金项目(编号：31201007)资助

From random mutagenesis to precise genome editing: the development and evolution of genome editing techniques in Drosophila

Fang Su¹, Zongliang Huang¹, Yawen Guo², Renjie Jiao^{1, 2}, Zi Li¹, Jianming Chen^{1, 3}, Jiyong Liu¹

1. Sino-French Hoffmann Institute, Guangzhou Medical University, Guangzhou 511436, China;
2. State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;
3. State Key Laboratory Breeding Base of Marine Genetic Resources, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China

Received:2015-07-27 Revised:2015-10-19 Online:2016-01-20 Published:2016-01-20
Supported by:
[Supported by the National Natural Science Foundation of China(No; 31201007)]

摘要/Abstract

摘要： 黑腹果蝇(Drosophila melanogaster)是研究生命科学的重要模式动物,基因组编辑技术的发展有助于人们更好地利用果蝇来进行遗传学、发育生物学、生物医学等领域的研究。从20世纪开始,果蝇基因组编辑技术经历了从随机突变到精确敲除、从单纯的基因突变体制作到多样化的基因组精确编辑的过程。甲基磺酸乙酯(Ethyl methanesulfonate, EMS)化学诱变是利用正向遗传学研究基因功能的重要手段,但是无法实现果蝇基因的精确敲除。基于同源重组建立的基因打靶技术首次实现对果蝇基因组任意位点的精确编辑,但效率较低。CRISPR/Cas9(Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein)系统介导的果蝇基因组精确编辑相对于基因打靶技术具有简单、快速、高效的特点。本文以果蝇基因敲除为主线,系统阐述了果蝇基因组编辑技术的演化,着重论述了基因打靶、ZFN(Zinc-finger nucleases)、TALEN(Transcription activator-like effector nucleases)及CRISPR/Cas9技术的发展和应用。

关键词: 果蝇, 基因组编辑, ZFN, TALEN, CRISPR/Cas9

Abstract: Drosophila melanogaster, an important model organism for studying life science, has contributed more to the research of genetics, developmental biology and biomedicine with the development of genome editing techniques. Drosophila genome-editing techniques have evolved from random mutagenesis to precise genome editing and from simple mutant construction to diverse genome editing methods since the 20th century. Chemical mutagenesis, using Ethyl methanesulfonate (EMS), is an important technique to study gene function in forward genetics, however, the precise knockout of Drosophila genes could not be achieved. The gene targeting technology, based on homologous recombination, has accomplished the precise editing of Drosophila genome for the first time, but with low efficiency. The CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein)-mediated precise genome editing is simple, fast and highly efficient compared with the gene targeting technology in Drosophila. In this review, we focus on Drosophila gene knockout, and summarize the evolution of genome editing techniques in Drosophila, emphasizing the development and applications of gene targeting, zinc-finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN) and CRISPR/Cas9 techniques.

Key words: Drosophila, genome editing, ZFN, TALEN, CRISPR/Cas9

苏方, 黄宗靓, 郭雅文, 焦仁杰, 李孜, 陈建明, 刘继勇. 从随机突变到精确编辑：果蝇基因组编辑技术的发展及演化[J]. 遗传, 2016, 38(1): 17-27.

Fang Su, Zongliang Huang, Yawen Guo, Renjie Jiao, Zi Li, Jianming Chen, Jiyong Liu. From random mutagenesis to precise genome editing: the development and evolution of genome editing techniques in Drosophila[J]. HEREDITAS(Beijing), 2016, 38(1): 17-27.

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从随机突变到精确编辑：果蝇基因组编辑技术的发展及演化

From random mutagenesis to precise genome editing: the development and evolution of genome editing techniques in Drosophila

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