Targeted gene disruption in somatic zebrafish cells using engineered TALENs

Targeted gene disruption in somatic zebrafish cells using engineered TALENs To the Editor: Miller et al . recently described a transcription activator–like effector nuclease (TALEN) architecture for efficient genome editing in cultured human cells. We sought to determine whether the same framework could be used to efficiently disrupt endogenous genes in somatic cells of zebrafish and how the efficiency of TALENs compares with that obtained using engineered zinc-finger nucleases (ZFNs). TALENs, which comprise an engineered array of transcription activator–like effector repeats fused to the nonspecific FokI cleavage domain, introduce targeted double-stranded breaks in human cells with high efficiency. Repair of these double-stranded breaks by normal DNA repair mechanisms, such as nonhomologous end-joining (NHEJ) or homologous recombination, enables introduction of alterations at or near the site of the break. A single 34-amino-acid transcription activator–like effector repeat binds to one bp of DNA, and repeats which bind each of the four DNA bases have been described . These modules can be assembled into arrays capable of binding extended DNA sequences. TALENs may have advantages over engineered ZFNs due to the relative ease with which they can be designed and their potential ability to be targeted to a wide range of sequences, with target sites reported to be as frequent as http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Biotechnology Springer Journals

Targeted gene disruption in somatic zebrafish cells using engineered TALENs

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Publisher
Springer Journals
Copyright
Copyright © 2011 Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.
ISSN
1087-0156
eISSN
1546-1696
D.O.I.
10.1038/nbt.1934
Publisher site
See Article on Publisher Site

Abstract

To the Editor: Miller et al . recently described a transcription activator–like effector nuclease (TALEN) architecture for efficient genome editing in cultured human cells. We sought to determine whether the same framework could be used to efficiently disrupt endogenous genes in somatic cells of zebrafish and how the efficiency of TALENs compares with that obtained using engineered zinc-finger nucleases (ZFNs). TALENs, which comprise an engineered array of transcription activator–like effector repeats fused to the nonspecific FokI cleavage domain, introduce targeted double-stranded breaks in human cells with high efficiency. Repair of these double-stranded breaks by normal DNA repair mechanisms, such as nonhomologous end-joining (NHEJ) or homologous recombination, enables introduction of alterations at or near the site of the break. A single 34-amino-acid transcription activator–like effector repeat binds to one bp of DNA, and repeats which bind each of the four DNA bases have been described . These modules can be assembled into arrays capable of binding extended DNA sequences. TALENs may have advantages over engineered ZFNs due to the relative ease with which they can be designed and their potential ability to be targeted to a wide range of sequences, with target sites reported to be as frequent as

Journal

Nature BiotechnologySpringer Journals

Published: Aug 5, 2011

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