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Methods DOI:10.1016/j.ymeth.2014.04.009

Methods for targeted mutagenesis in zebrafish using TALENs.

Publication TypeJournal Article
Year of Publication2014
AuthorsHwang, WY, Peterson, RT, Yeh, J-RJoanna
JournalMethods
Volume69
Issue1
Pages76-84
Date Published2014 Aug 15
ISSN1095-9130
KeywordsAnimals, Deoxyribonucleases, Mutagenesis, Site-Directed, Protein Engineering, Zebrafish
Abstract

The transcription activator-like effector (TALE) nucleases, or TALENs, are customizable restriction enzymes that may be used to induce mutations at nearly any investigator-specified DNA sequence in zebrafish. The DNA-binding specificities of TALENs are determined by a protein array comprised of four types of TALE repeats, where each repeat recognizes a different DNA base. Here, we describe methods for constructing TALEN vectors that have been shown to achieve high success rates and mutation efficiencies in zebrafish. In addition, we discuss simple techniques and protocols that can be used to detect TALEN-induced mutations at almost any genomic locus. These methods should enable zebrafish researchers to quickly generate targeted mutations at their genes-of-interest.

URLhttp://linkinghub.elsevier.com/retrieve/pii/S1046-2023(14)00153-4
DOI10.1016/j.ymeth.2014.04.009
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/24747922?dopt=Abstract

Alternate JournalMethods
PubMed ID24747922
PubMed Central IDPMC4157959
Grant ListR01 CA140188 / CA / NCI NIH HHS / United States
R01 GM088040 / GM / NIGMS NIH HHS / United States