Electroporation enables the efficient mRNA delivery into the mouse zygotes and facilitates CRISPR/Cas9-based genome editing

被引：224

作者：

Hashimoto, Masakazu ^{[1
]}

Takemoto, Tatsuya ^{[2
]}

机构：

[1] Chiba Univ, Grad Sch Med, Div Dev Biol, Chuo Ku, Chiba 2608670, Japan

[2] Univ Tokushima, Fujii Mem Inst Med Sci, Div Embryol, Tokushima 7708503, Japan

来源：

SCIENTIFIC REPORTS | 2015年 / 5卷

关键词：

ONE-STEP GENERATION; MICE; REPORTER; EMBRYOS; TALEN;

D O I：

10.1038/srep11315

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：

Recent use of the CRISPR/Cas9 system has dramatically reduced the time required to produce mutant mice, but the involvement of a time-consuming microinjection step still hampers its application for high-throughput genetic analysis. Here we developed a simple, highly efficient, and large-scale genome editing method, in which the RNAs for the CRISPR/Cas9 system are electroporated into zygotes rather than microinjected. We used this method to perform single-stranded oligodeoxynucleotide (ssODN)-mediated knock-in in mouse embryos. This method facilitates large-scale genetic analysis in the mouse.

引用

页数：7

共 10 条

[1] Establishment of Conditional Reporter Mouse Lines at ROSA26 Locus For Live Cell Imaging [J].

Abe, Takaya ;

Kiyonari, Hiroshi ;

Shioi, Go ;

Inoue, Ken-Ichi ;

Nakao, Kazuki ;

Aizawa, Shinichi ;

Fujimori, Toshihiko .

GENESIS, 2011, 49 (07) :579-590

[2] 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

[3] 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

[4] Efficient delivery of dsRNA into zona-enclosed mouse oocytes and preimplantation embryos by electroporation [J].

Grabarek, JB ;

Plusa, B ;

Glover, DM ;

Zernicka-Goetz, M .

GENESIS, 2002, 32 (04) :269-276

[5] Simple knockout by electroporation of engineered endonucleases into intact rat embryos [J].

Kaneko, Takehito ;

Sakuma, Tetsushi ;

Yamamoto, Takashi ;

Mashimo, Tomoji .

SCIENTIFIC REPORTS, 2014, 4

[6] Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9 and single guided RNA [J].

Mashiko, Daisuke ;

Fujihara, Yoshitaka ;

Satouh, Yuhkoh ;

Miyata, Haruhiko ;

Isotani, Ayako ;

Ikawa, Masahito .

SCIENTIFIC REPORTS, 2013, 3

[7] Fgf10 is essential for limb and lung formation [J].

Sekine, K ;

Ohuchi, H ;

Fujiwara, M ;

Yamasaki, M ;

Yoshizawa, T ;

Sato, T ;

Yagishita, N ;

Matsui, D ;

Koga, Y ;

Itoh, N ;

Kato, S .

NATURE GENETICS, 1999, 21 (01) :138-141

[8] One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering [J].

Wang, Haoyi ;

Yang, Hui ;

Shivalila, Chikdu S. ;

Dawlaty, Meelad M. ;

Cheng, Albert W. ;

Zhang, Feng ;

Jaenisch, Rudolf .

CELL, 2013, 153 (04) :910-918

[9] One-Step Generation of Mice Carrying Reporter and Conditional Alleles by CRISPR/Cas-Mediated Genome Engineering [J].

Yang, Hui ;

Wang, Haoyi ;

Shivalila, Chikdu S. ;

Cheng, Albert W. ;

Shi, Linyu ;

Jaenisch, Rudolf .

CELL, 2013, 154 (06) :1370-1379

[10] Highly efficient targeted mutagenesis in one-cell mouse embryos mediated by the TALEN and CRISPR/Cas systems [J].

Yasue, Akihiro ;

Mitsui, Silvia Naomi ;

Watanabe, Takahito ;

Sakuma, Tetsushi ;

Oyadomari, Seiichi ;

Yamamoto, Takashi ;

Noji, Sumihare ;

Mito, Taro ;

Tanaka, Eiji .

SCIENTIFIC REPORTS, 2014, 4

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