Year: 2019
Type: preprint
Abstract: 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.
Source: Genome Research
Authors: Yonghua Sun, Bo Zhang, Lingfei Luo, De-Li Shi, Han Wang, Zongbin Cui, Honghui Huang, Ying Cao, Xiaodong Shu, Wenqing Zhang, Jianfeng Zhou, Yun Li, Jiulin Du, Qingshun Zhao, Jun Chen, Hanbing Zhong, Tao P. Zhong, Li Li, Jing-Wei Xiong, Jinrong Peng, Wuhan Xiao, Jian Zhang, Jihua Yao, Zhan Yin, Xianming Mo, Gang Peng, Jun Zhu, Yan Chen, Yong Zhou, Dong Liu, Weijun Pan, Yiyue Zhang, Hua Ruan, Feng Liu, Zuoyan Zhu, Anming Meng
Institutions: Chinese Academy of Sciences, Institute of Hydrobiology, State Key Laboratory of Freshwater Ecology and Biotechnology, Peking University, Southwest University, Guangdong Medical College, Soochow University, Tongji University, Guangzhou Institutes of Biomedicine and Health, South China University of Technology, Ocean University of China, State Key Laboratory of Neuroscience, Model Animal Research Center, Nanjing University, Zhejiang University, Southern University of Science and Technology, East China Normal University, Yunnan University, Fudan University, Sichuan University, West China Hospital of Sichuan University, State Key Laboratory of Biotherapy, Shanghai Jiao Tong University, Ruijin Hospital, Shanghai Institute of Nutrition and Health, Institute of Zoology, State Key Laboratory of Membrane Biology, Tsinghua University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2994882832