Programmable base editing of zebrafish genome using a modified CRISPR-Cas9 system

被引:174
作者
Zhang, Yihan [1 ,2 ]
Qin, Wei [1 ]
Lu, Xiaochan [1 ]
Xu, Jason [2 ]
Huang, Haigen [2 ]
Bai, Haipeng [1 ]
Li, Song [1 ]
Lin, Shuo [1 ,2 ]
机构
[1] Peking Univ, Shenzhen Grad Sch, Sch Chem Biol & Biotechnol, Lab Chem Genom, Shenzhen 518055, Peoples R China
[2] Univ Calif Los Angeles, Dept Mol Cell & Dev Biol, Los Angeles, CA 90095 USA
关键词
PRECISE; VARIANTS; RICE; GENE; DNA;
D O I
10.1038/s41467-017-00175-6
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Precise genetic modifications in model animals are essential for biomedical research. Here, we report a programmable "base editing" system to induce precise base conversion with high efficiency in zebrafish. Using cytidine deaminase fused to Cas9 nickase, up to 28% of site-specific single-base mutations are achieved in multiple gene loci. In addition, an engineered Cas9-VQR variant with 5'-NGA PAM specificities is used to induce base conversion in zebrafish. This shows that Cas9 variants can be used to expand the utility of this technology. Collectively, the targeted base editing system represents a strategy for precise and effective genome editing in zebrafish.
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页数:5
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共 23 条
[1]   Genome editing using CRISPR/Cas9-based knock-in approaches in zebrafish [J].
Albadri, Shahad ;
Del Bene, Filippo ;
Revenu, Celine .
METHODS, 2017, 121 :77-85
[2]   Highly efficient CRISPR/Cas9-mediated knock-in in zebrafish by homology-independent DNA repair [J].
Auer, Thomas O. ;
Duroure, Karine ;
De Cian, Anne ;
Concordet, Jean-Paul ;
Del Bene, Filippo .
GENOME RESEARCH, 2014, 24 (01) :142-153
[3]   Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 System [J].
Bassett, Andrew R. ;
Tibbit, Charlotte ;
Ponting, Chris P. ;
Liu, Ji-Long .
CELL REPORTS, 2013, 4 (01) :220-228
[4]   Evolution of Phosphorylation-Dependent Regulation of Activation-Induced Cytidine Deaminase [J].
Basu, Uttiya ;
Wang, Yabin ;
Alt, Frederick W. .
MOLECULAR CELL, 2008, 32 (02) :285-291
[5]   In vivo genome editing using a high-efficiency TALEN system [J].
Bedell, Victoria M. ;
Wang, Ying ;
Campbell, Jarryd M. ;
Poshusta, Tanya L. ;
Starker, Colby G. ;
Krug, Randall G., II ;
Tan, Wenfang ;
Penheiter, Sumedha G. ;
Ma, Alvin C. ;
Leung, Anskar Y. H. ;
Fahrenkrug, Scott C. ;
Carlson, Daniel F. ;
Voytas, Daniel F. ;
Clark, Karl J. ;
Essner, Jeffrey J. ;
Ekker, Stephen C. .
NATURE, 2012, 491 (7422) :114-U133
[6]   Cas9 Variants Expand the Target Repertoire in Caenorhabditis elegans [J].
Bell, Ryan T. ;
Fu, Becky X. H. ;
Fire, Andrew Z. .
GENETICS, 2016, 202 (02) :381-+
[7]   Enhancing gene targeting with designed zinc finger nucleases [J].
Bibikova, M ;
Beumer, K ;
Trautman, JK ;
Carroll, D .
SCIENCE, 2003, 300 (5620) :764-764
[8]   Multiplex Genome Engineering Using CRISPR/Cas Systems [J].
Cong, Le ;
Ran, F. Ann ;
Cox, David ;
Lin, Shuailiang ;
Barretto, Robert ;
Habib, Naomi ;
Hsu, Patrick D. ;
Wu, Xuebing ;
Jiang, Wenyan ;
Marraffini, Luciano A. ;
Zhang, Feng .
SCIENCE, 2013, 339 (6121) :819-823
[9]   Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9 [J].
Feng, Yan ;
Chen, Cheng ;
Han, Yuxiang ;
Chen, Zelin ;
Lu, Xiaochan ;
Liang, Fang ;
Li, Song ;
Qin, Wei ;
Lin, Shuo .
G3-GENES GENOMES GENETICS, 2016, 6 (08) :2517-2521
[10]   Efficient genome editing in zebrafish using a CRISPR-Cas system [J].
Hwang, Woong Y. ;
Fu, Yanfang ;
Reyon, Deepak ;
Maeder, Morgan L. ;
Tsai, Shengdar Q. ;
Sander, Jeffry D. ;
Peterson, Randall T. ;
Yeh, J-R Joanna ;
Joung, J. Keith .
NATURE BIOTECHNOLOGY, 2013, 31 (03) :227-229