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PMID: 21179091 Published · ppublish English Journal Article

A TALE nuclease architecture for efficient genome editing.

Nature biotechnology ·Vol. 29 ·No. 2 ·2011-02-00 ·Pages 143-8

Miller JC, Tan S, Qiao G, Barlow KA, Wang J, Xia DF, Meng X, Paschon DE, Leung E, Hinkley SJ, Dulay GP, Hua KL, Ankoudinova I, Cost GJ, Urnov FD, Zhang HS, Holmes MC, Zhang L, Gregory PD, Rebar EJ

Abstract

Nucleases that cleave unique genomic sequences in living cells can be used for targeted gene editing and mutagenesis. Here we develop a strategy for generating such reagents based on transcription activator-like effector (TALE) proteins from Xanthomonas. We identify TALE truncation variants that efficiently cleave DNA when linked to the catalytic domain of FokI and use these nucleases to generate discrete edits or small deletions within endogenous human NTF3 and CCR5 genes at efficiencies of up to 25%. We further show that designed TALEs can regulate endogenous mammalian genes. These studies demonstrate the effective application of designed TALE transcription factors and nucleases for the targeted regulation and modification of endogenous genes.

MeSH Terms
Bacterial Proteins/genetics,metabolism Base Sequence Binding Sites Combinatorial Chemistry Techniques/methods DNA/genetics,metabolism Deoxyribonucleases, Type II Site-Specific/genetics,metabolism Genetic Engineering Genome Humans K562 Cells Molecular Sequence Data Mutagenesis, Site-Directed/methods Receptors, CCR5/genetics Transcription Factors/genetics,metabolism Vascular Endothelial Growth Factor A/genetics Xanthomonas
Chemicals
Bacterial Proteins Receptors, CCR5 Transcription Factors VEGFA protein, human Vascular Endothelial Growth Factor A DNA endodeoxyribonuclease FokI Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
20 authors, click to expand affiliations / ORCID
Miller Jeffrey C
Sangamo BioSciences, Inc., Richmond, California, USA.
Tan Siyuan
Qiao Guijuan
Barlow Kyle A
Wang Jianbin
Xia Danny F
Meng Xiangdong
Paschon David E
Leung Elo
Hinkley Sarah J
Dulay Gladys P
Hua Kevin L
Ankoudinova Irina
Cost Gregory J
Urnov Fyodor D
Zhang H Steve
Holmes Michael C
Zhang Lei
Gregory Philip D
Rebar Edward J
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Article Info
Journal
Nature biotechnology
Abbr.
Nat Biotechnol
ISSN
1546-1696
DOI
10.1038/nbt.1755
Published
2011-02-00
Epub
2010-00-22
Pages
143-8
Language
English
Region
United States
NLM ID
9604648
Subset
IM
Corrections
CommentIn
PMID: 21301438
CommentIn
PMID: 21473016
CommentIn
PMID: 21822240
CommentIn
PMID: 21822241
CommentIn
PMID: 21822242
External Links
PubMed source DOI: 10.1038/nbt.1755 View journal details
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