RNA-Guided Genome Editing in Drosophila with the Purified Cas9 Protein

被引:33
作者
Lee, Jeong-Soo [1 ,2 ]
Kwak, Su-Jin [1 ]
Kim, Jungeun [3 ,4 ]
Kim, Ae-Kyeong [1 ]
Noh, Hae Min [1 ]
Kim, Jin-Soo [3 ,4 ,5 ]
Yu, Kweon [1 ,2 ]
机构
[1] KRIBB, Neurophysiol Res Grp, Bionano Ctr, Taejon 305806, South Korea
[2] KUST, Funct Genom Dept, Taejon 305333, South Korea
[3] Seoul Natl Univ, Natl Creat Res Initiat Ctr Genome Engn, Seoul 151747, South Korea
[4] Seoul Natl Univ, Dept Chem, Seoul 151747, South Korea
[5] Inst for Basic Sci, Ctr Genome Engn, Taejon, South Korea
关键词
Cas9; protein; RGEN; Drosophila; GENE KNOCKOUT; HUMAN-CELLS; ENDONUCLEASES; NUCLEASES; CRISPR/CAS9; MICE;
D O I
10.1534/g3.114.012179
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
We report a method for generating Drosophila germline mutants effectively via injection of the complex of the purified Cas9 protein, tracrRNA, and gene-specific crRNAs, which may reduce delayed mutations because of the transient activity of the Cas9 protein, combined with the simple mutation detection in GO founders by the T7E1 assay.
引用
收藏
页码:1291 / 1295
页数:5
相关论文
共 25 条
[1]   Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases [J].
Bae, Sangsu ;
Park, Jeongbin ;
Kim, Jin-Soo .
BIOINFORMATICS, 2014, 30 (10) :1473-1475
[2]   CRISPR/Cas9 mediated genome engineering in Drosophila [J].
Bassett, Andrew ;
Liu, Ji-Long .
METHODS, 2014, 69 (02) :128-136
[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]   Targeted genome engineering techniques in Drosophila [J].
Beumer, Kelly J. ;
Carroll, Dana .
METHODS, 2014, 68 (01) :29-37
[5]   Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases [J].
Cho, Seung Woo ;
Kim, Sojung ;
Kim, Yongsub ;
Kweon, Jiyeon ;
Kim, Heon Seok ;
Bae, Sangsu ;
Kim, Jin-Soo .
GENOME RESEARCH, 2014, 24 (01) :132-141
[6]   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-+
[7]   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
[8]   ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering [J].
Gaj, Thomas ;
Gersbach, Charles A. ;
Barbas, Carlos F., III .
TRENDS IN BIOTECHNOLOGY, 2013, 31 (07) :397-405
[9]   CRISPR/Cas9-mediated genome engineering and the promise of designer flies on demand [J].
Gratz, Scott J. ;
Wildonger, Jill ;
Harrison, Melissa M. ;
O'Connor-Giles, Kate M. .
FLY, 2013, 7 (04) :249-255
[10]   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-+