Efficient targeted gene disruption in Xenopus embryos using engineered transcription activator-like effector nucleases (TALENs)

被引：192

作者：

Lei, Yong ^{[1
]}

Guo, Xiaogang ^{[2
]}

Liu, Yun ^{[1
]}

Cao, Yang ^{[2
]}

Deng, Yi ^{[3
]}

Chen, Xiongfeng ^{[4
]}

Cheng, Christopher H. K. ^{[1
,5
]}

Dawid, Igor B. ^{[6
]}

Chen, Yonglong ^{[2
]}

Zhao, Hui ^{[1
,5
]}

机构：

[1] Chinese Univ Hong Kong, Fac Med, Sch Biomed Sci, Shatin, Hong Kong, Peoples R China

[2] Chinese Acad Sci, Guangzhou Inst Biomed & Hlth, Key Lab Regenerat Biol, Guangzhou 510530, Guangdong, Peoples R China

[3] S Univ Sci & Technol China, Dept Biol, Shenzhen 518055, Peoples R China

[4] NCI, Adv Biomed Comp Ctr, NIH, Frederick, MD 21702 USA

[5] Chinese Univ Hong Kong, Shenzhen Res Inst, Shenzhen 518057, Peoples R China

[6] Eunice Kennedy Shriver Natl Inst Child Hlth & Hum, Program Genom Differentiat, NIH, Bethesda, MD 20892 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2012年 / 109卷 / 43期

基金：

美国国家卫生研究院; 中国国家自然科学基金;

关键词：

genome editing; heritable mutagenesis; mutagenesis detection; reverse genetics; genome engineering; DNA-BINDING SPECIFICITY; III EFFECTORS; RESTRICTION ENZYMES; FINGER; CLEAVAGE; RECOGNITION; PANCREAS; EXOCRINE; BREAKING; CODE;

D O I：

10.1073/pnas.1215421109

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：

Transcription activator-like effector nucleases (TALENs) are an approach for directed gene disruption and have been proved to be effective in various animal models. Here, we report that TALENs can induce somatic mutations in Xenopus embryos with reliably high efficiency and that such mutations are heritable through germ-line transmission. We modified the Golden Gate method for TALEN assembly to make the product suitable for RNA transcription and microinjection into Xenopus embryos. Eight pairs of TALENs were constructed to target eight Xenopus genes, and all resulted in indel mutations with high efficiencies of up to 95.7% at the targeted loci. Furthermore, mutations induced by TALENs were highly efficiently passed through the germ line to F-1 frogs. Together with simple and reliable PCR-based approaches for detecting TALEN-induced mutations, our results indicate that TALENs are an effective tool for targeted gene editing/knockout in Xenopus.

引用

页码：17484 / 17489

页数：6

共 24 条

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