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
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.