Heritable Gene Knockout in Caenorhabditis elegans by Direct Injection of Cas9-sgRNA Ribonucleoproteins

被引:208
作者
Cho, Seung Woo [1 ]
Lee, Jihyun [2 ]
Carroll, Dana [4 ]
Kim, Jin-Soo [1 ]
Lee, Junho [2 ,3 ]
机构
[1] Seoul Natl Univ, Dept Chem, Seoul 151742, South Korea
[2] Seoul Natl Univ, Dept Biol Sci, Seoul 151742, South Korea
[3] Seoul Natl Univ, Dept Biophys & Chem Biol, Seoul 151742, South Korea
[4] Univ Utah, Sch Med, Dept Biochem, Salt Lake City, UT 84112 USA
基金
美国国家卫生研究院; 新加坡国家研究基金会;
关键词
Caenorhabditis elegans; Cas9-sgRNA ribonucleoprotein; NHEJ; genome editing; HUMAN-CELLS; GENOME; UNC-1;
D O I
10.1534/genetics.113.155853
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
We present a novel method of targeted gene disruption that involves direct injection of recombinant Cas9 protein complexed with guide RNA into the gonad of the nematode Caenorhabditis elegans. Biallelic mutants were recovered among the F-1 progeny, demonstrating the high efficiency of this method.
引用
收藏
页码:1177 / +
页数:5
相关论文
共 24 条
[1]  
BLAXTER ML, 1993, J BIOL CHEM, V268, P6600
[2]   UNC-1 regulates gap junctions important to locomotion in C. elegans [J].
Chen, Bojun ;
Liu, Qiang ;
Ge, Qian ;
Xie, Jia ;
Wang, Zhao-Wen .
CURRENT BIOLOGY, 2007, 17 (15) :1334-1339
[3]   Transgene-Free Genome Editing in Caenorhabditis elegans Using CRISPR-Cas [J].
Chiu, Hui ;
Schwartz, Hillel T. ;
Antoshechkin, Igor ;
Sternberg, Paul W. .
GENETICS, 2013, 195 (03) :1167-1171
[4]   Heritable Gene Knockout in Caenorhabditis elegans by Direct Injection of Cas9-sgRNA Ribonucleoproteins [J].
Cho, Seung Woo ;
Lee, Jihyun ;
Carroll, Dana ;
Kim, Jin-Soo ;
Lee, Junho .
GENETICS, 2013, 195 (03) :1177-+
[5]   Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease [J].
Cho, Seung Woo ;
Kim, Sojung ;
Kim, Jong Min ;
Kim, Jin-Soo .
NATURE BIOTECHNOLOGY, 2013, 31 (03) :230-232
[6]   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
[7]   Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems [J].
DiCarlo, James E. ;
Norville, Julie E. ;
Mali, Prashant ;
Rios, Xavier ;
Aach, John ;
Church, George M. .
NUCLEIC ACIDS RESEARCH, 2013, 41 (07) :4336-4343
[8]   Heritable genome editing in C. elegans via a CRISPR-Cas9 system [J].
Friedland, Ari E. ;
Tzur, Yonatan B. ;
Esvelt, Kevin M. ;
Colaiacovo, Monica P. ;
Church, George M. ;
Calarco, John A. .
NATURE METHODS, 2013, 10 (08) :741-+
[9]   Genome Engineering of Drosophila with the CRISPR RNA-Guided Cas9 Nuclease [J].
Gratz, Scott J. ;
Cummings, Alexander M. ;
Nguyen, Jennifer N. ;
Hamm, Danielle C. ;
Donohue, Laura K. ;
Harrison, Melissa M. ;
Wildonger, Jill ;
O'Connor-Giles, Kate M. .
GENETICS, 2013, 194 (04) :1029-+
[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