PUBLICATION

A protocol for TALEN construction and gene targeting in zebrafish

Authors
Shen, Y., Huang, P., and Zhang, B.
ID
ZDB-PUB-130610-35
Date
2013
Source
Yi chuan = Hereditas   35(4): 533-544 (Journal)
Registered Authors
Shen, Yan, Zhang, Bo
Keywords
none
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Endonucleases/chemistry
  • Endonucleases/genetics*
  • Endonucleases/isolation & purification
  • Endonucleases/metabolism
  • Gene Targeting/methods*
  • Genetic Vectors/genetics
  • Molecular Sequence Data
  • Protein Engineering/methods*
  • Restriction Mapping
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed
23659945 Full text @ Yi Chuan
Abstract

TALEN (Transcription activator-like effector nuclease) is a newly developed family of artificially engineered sequence-specific endonucleases, which consists of a highly specific and repetitive DNA-binding domain derived from TALE (Transcription activator-like effector) and fused with the non-specific endonuclease domain of Fok`. TALENs have been reported to be able to induce site specific genome modification in quite a few species and in vitro cultured cells. Here, we introduced the principles for target site selection and confirmation and described a brief experimental protocol for the easy construction of customized TALENs using unit assembly (UA) method and also for generation of and screening for TALEN-mediated targeted zebrafish mutants through microinjection of TALEN mRNAs into zebrafish embryos. Theoretically, the principles and methods we described here are also applicable to gene targeting in other species.

Errata / Notes
Article is in Chinese.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping