Heritable and Precise Zebrafish Genome Editing Using a CRISPR-Cas System

被引：264

作者：

Hwang, Woong Y. ^{[1
,2
]}

Fu, Yanfang ^{[3
,4
,5
]}

Reyon, Deepak ^{[3
,4
,5
]}

Maeder, Morgan L. ^{[3
,4
]}

Kaini, Prakriti ^{[1
,2
]}

Sander, Jeffry D. ^{[3
,4
,5
]}

Joung, J. Keith ^{[3
,4
,5
]}

Peterson, Randall T. ^{[1
,2
,6
]}

Yeh, Jing-Ruey Joanna ^{[1
,2
]}

机构：

[1] Massachusetts Gen Hosp, Cardiovasc Res Ctr, Charlestown, MA 02129 USA

[2] Harvard Univ, Sch Med, Dept Med, Boston, MA USA

[3] Massachusetts Gen Hosp, Ctr Canc Res, Mol Pathol Unit, Charlestown, MA USA

[4] Massachusetts Gen Hosp, Ctr Computat & Integrat Biol, Charlestown, MA USA

[5] Harvard Univ, Sch Med, Dept Pathol, Boston, MA 02115 USA

[6] Broad Inst, Cambridge, MA USA

来源：

PLOS ONE | 2013年 / 8卷 / 07期

基金：

美国国家卫生研究院;

关键词：

ZINC-FINGER NUCLEASES; TARGETED GENE DISRUPTION; TALENS; ENDONUCLEASE; MUTAGENESIS; BACTERIA; ARCHAEA; CELLS;

D O I：

10.1371/journal.pone.0068708

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：

We have previously reported a simple and customizable CRISPR (clustered regularly interspaced short palindromic repeats) RNA-guided Cas9 nuclease (RGN) system that can be used to efficiently and robustly introduce somatic indel mutations in endogenous zebrafish genes. Here we demonstrate that RGN-induced mutations are heritable, with efficiencies of germline transmission reaching as high as 100%. In addition, we extend the power of the RGN system by showing that these nucleases can be used with single-stranded oligodeoxynucleotides (ssODNs) to create precise intended sequence modifications, including single nucleotide substitutions. Finally, we describe and validate simple strategies that improve the targeting range of RGNs from 1 in every 128 basepairs (bps) of random DNA sequence to 1 in every 8 bps. Together, these advances expand the utility of the CRISPR-Cas system in the zebrafish beyond somatic indel formation to heritable and precise genome modifications.

引用

页数：9

共 35 条

[1]

[Anonymous], GENOME RES

[2] 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

[3] The CRISPR System-Keeping Zebrafish Gene Targeting Fresh [J].

Blackburn, Patrick R. ;

Campbell, Jarryd M. ;

Clark, Karl J. ;

Ekker, Stephen C. .

ZEBRAFISH, 2013, 10 (01) :116-118

[4] Highly efficient generation of heritable zebrafish gene mutations using homo- and heterodimeric TALENs [J].

Cade, Lindsay ;

Reyon, Deepak ;

Hwang, Woong Y. ;

Tsai, Shengdar Q. ;

Patel, Samir ;

Khayter, Cyd ;

Joung, J. Keith ;

Sander, Jeffry D. ;

Peterson, Randall T. ;

Yeh, Jing-Ruey Joanna .

NUCLEIC ACIDS RESEARCH, 2012, 40 (16) :8001-8010

[5] Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting [J].

Cermak, Tomas ;

Doyle, Erin L. ;

Christian, Michelle ;

Wang, Li ;

Zhang, Yong ;

Schmidt, Clarice ;

Baller, Joshua A. ;

Somia, Nikunj V. ;

Bogdanove, Adam J. ;

Voytas, Daniel F. .

NUCLEIC ACIDS RESEARCH, 2011, 39 (12) :e82

[6] Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos [J].

Chang, Nannan ;

Sun, Changhong ;

Gao, Lu ;

Zhu, Dan ;

Xu, Xiufei ;

Zhu, Xiaojun ;

Xiong, Jing-Wei ;

Xi, Jianzhong Jeff .

CELL RESEARCH, 2013, 23 (04) :465-472

[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] 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] Simple Methods for Generating and Detecting Locus-Specific Mutations Induced with TALENs in the Zebrafish Genome [J].

Dahlem, Timothy J. ;

Hoshijima, Kazuyuki ;

Jurynec, Michael J. ;

Gunther, Derrick ;

Starker, Colby G. ;

Locke, Alexandra S. ;

Weis, Allison M. ;

Voytas, Daniel F. ;

Grunwald, David Jonah .

PLOS GENETICS, 2012, 8 (08)

[10] 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

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