Document Detail


Efficient targeted gene disruption in Xenopus embryos using engineered transcription activator-like effector nucleases (TALENs).
MedLine Citation:
PMID:  23045671     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Yong Lei; Xiaogang Guo; Yun Liu; Yang Cao; Yi Deng; Xiongfeng Chen; Christopher H K Cheng; Igor B Dawid; Yonglong Chen; Hui Zhao
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-10-08
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  109     ISSN:  1091-6490     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-24     Completed Date:  2013-01-07     Revised Date:  2013-07-11    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  17484-9     Citation Subset:  IM    
Affiliation:
School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People's Republic of China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
DNA
Embryo, Nonmammalian
Endonucleases / metabolism*
Gene Targeting*
Molecular Sequence Data
Trans-Activators / metabolism*
Xenopus / embryology*,  genetics
Chemical
Reg. No./Substance:
0/Trans-Activators; 9007-49-2/DNA; EC 3.1.-/Endonucleases
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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