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ZFIN ID: ZDB-PUB-140513-116
Methods for targeted mutagenesis in zebrafish using TALENs
Hwang, W.Y., Peterson, R.T., Yeh, J.R.
Date: 2014
Source: Methods (San Diego, Calif.)   69(1): 76-84 (Journal)
Registered Authors: Peterson, Randall, Yeh, Jing-Ruey (Joanna)
Keywords: Gene disruption, Gene targeting, Genome engineering, Knockout, Site-specific nuclease, Transcription activator-like effector nuclease, Zebrafish
MeSH Terms:
  • Animals
  • Deoxyribonucleases/chemistry
  • Deoxyribonucleases/genetics
  • Mutagenesis, Site-Directed/methods*
  • Protein Engineering
  • Zebrafish/genetics*
PubMed: 24747922 Full text @ Methods
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.