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Systematic genome editing of the genes on zebrafish Chromosome 1 by CRISPR/Cas9.

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作者：
Yonghua Sun[1] * ; Bo Zhang[2] ; Lingfei Luo[3] ; De-Li Shi[4] ; Han Wang[5] ; Zongbin Cui[1] ; Honghui Huang[3] ; Ying Cao[6] ; Xiaodong Shu[7] ; Wenqing Zhang[8] ; Jianfeng Zhou[9] ; Yun Li[9] ; Jiulin Du[10] ; Qingshun Zhao[11] ; Jun Chen[12] ; Hanbing Zhong[13] ; Tao P. Zhong[14] ; Li Li[3] ; Jing-Wei Xiong[15] ; Jinrong Peng[12] ; Wuhan Xiao[1] ; Jian Zhang[16] ; Jihua Yao[17] ; Zhan Yin[1] ; Xianming Mo[18] ; Gang Peng[19] ; Jun Zhu[20] ; Yan Chen[21] ; Yong Zhou[22] ; Dong Liu[13] ; Weijun Pan[22] ; Yiyue Zhang[8] ; Hua Ruan[3] ; Feng Liu[23] ; Zuoyan Zhu[1] ; Anming Meng[24] ; The ZAKOC Consortium ;
机构： [1]State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Innovation Academy for Seed Design, Chinese Academy of Sciences, Wuhan, Hubei, 430072, China [2]Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, Peking University Genome Editing Research Center, College of Life Sciences, Peking University, Beijing, 100871, China [3]School of Life Sciences, Southwest University, Chongqing, 400715, China [4]Guangdong Medical University, Zhanjiang, Guangdong, 524023, China [5]Center for Circadian Clocks, Soochow University, Suzhou, Jiangsu, 215123, China [6]School of Life Sciences and Technology, Tongji University, Shanghai, 200092, China [7]Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, Guangdong, 510530, China [8]Division of Cell, Developmental and Integrative Biology, School of Medicine, South China University of Technology, Guangzhou, Guangdong, 510006, China [9]School of Medicine and Pharmacy, Ocean University of China, Qingdao, Shandong, 266100, China [10]State Key Laboratory of Neuroscience, Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, 200031, China [11]Model Animal Research Center, Nanjing University, Nanjing, Jiangsu, 210093, China [12]College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, China [13]Department of Biology, South University of Science and Technology of China, Shenzhen, Guangdong, 518055, China [14]Institute of Biomedical Sciences, East China Normal University, Shanghai, 200062, China [15]College of Life Sciences, Institute of Molecular Medicine, Peking University, Beijing, 100871, China [16]School of Life Sciences, Yunnan University, Kunming, Yunnan, 650091, China [17]School of Life Sciences, Fudan University, Shanghai, 200433, China [18]State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, China [19]Institutes of Brain Science, Fudan University, Shanghai, 200433, China [20]Sino- French Research Center for Life Sciences and Genomics, Rui-Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China [21]Institute of Health Sciences, Chinese Academy of Sciences & Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China [22]CAS Key Laboratory of Nutrition, Metabolism and Food Safety, Shanghai Institute of Nutrition and Health, Chinese Academy of Sciences, Shanghai, 200031, China [23]State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China [24]School of Life Sciences, Tsinghua University, Beijing, 100084, China
出处：
DOI： 10.1101/gr.248559.119
ISSN：

摘要：
Genome editing by the well-established CRISPR/Cas9 technology has greatly facilitated our understanding of many biological processes. However, a complete whole-genome knockout for any species or model organism has rarely been achieved. Here, we performed a systematic knockout of all the genes (1333) on Chromosome 1 in zebrafish, successfully mutated 1029 genes, and generated 1039 germline-transmissible alleles corresponding to 636 genes. Meanwhile, by high-throughput bioinformatics analysis, we found that sequence features play pivotal roles in effective gRNA targeting at specific genes of interest, while the success rate of gene targeting positively correlates with GC content of the target sites. Moreover, we found that nearly one-fourth of all mutants are related to human diseases, and several representative CRISPR/Cas9-generated mutants are described here. Furthermore, we tried to identify the underlying mechanisms leading to distinct phenotypes between genetic mutants and antisense morpholino-mediated knockdown embryos. Altogether, this work has generated the first chromosome-wide collection of zebrafish genetic mutants by the CRISPR/Cas9 technology, which will serve as a valuable resource for the community, and our bioinformatics analysis also provides some useful guidance to design gene-specific gRNAs for successful gene editing. © 2020 Sun et al.; Published by Cold Spring Harbor Laboratory Press.

基金：
This work was supported by grants from the Ministry of Science and Technology of China (2018YFA0801000, 2018YFA0800200), the National Natural Science Foundation of China (31830061, 31721005), and the Strategic Priority Research Program of the Chinese Academy of Sciences, China.
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外文
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出版当年[2019]版：
大类 | 1 区 生物
小类 | 1 区 生化与分子生物学 1 区 生物工程与应用微生物 1 区 遗传学
最新[2023]版：
大类 | 2 区 生物学
小类 | 1 区 生物工程与应用微生物 2 区 生化与分子生物学 2 区 遗传学
第一作者：
第一作者机构： [1]State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Innovation Academy for Seed Design, Chinese Academy of Sciences, Wuhan, Hubei, 430072, China
通讯作者：
推荐引用方式(GB/T 7714)：
Yonghua Sun,Bo Zhang,Lingfei Luo,et al.Systematic genome editing of the genes on zebrafish Chromosome 1 by CRISPR/Cas9.[J].Genome research.2019,doi:10.1101/gr.248559.119.
APA：
Yonghua Sun,Bo Zhang,Lingfei Luo,De-Li Shi,Han Wang...&The ZAKOC Consortium.(2019).Systematic genome editing of the genes on zebrafish Chromosome 1 by CRISPR/Cas9..Genome research,,
MLA：
Yonghua Sun,et al."Systematic genome editing of the genes on zebrafish Chromosome 1 by CRISPR/Cas9.".Genome research .(2019)

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