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Updated: May 15, 2026
12:04
Mouse Genome Engineering Using Designer Nucleases
Published on: April 2, 2014
Efficient targeted mutagenesis in medaka using custom-designed transcription activator-like effector nucleases
Satoshi Ansai1, Tetsushi Sakuma, Takashi Yamamoto
1Division of Applied Bioscience, Graduate School of Agriculture Kyoto University, Sakyo-ku, Kyoto, Japan.
Genetics
|January 5, 2013
Summary
Transcription activator-like effector nucleases (TALENs) enable efficient targeted genome editing in medaka fish. This study establishes TALENs for rapid generation of DJ-1 gene knockout medaka, advancing their use as a vertebrate model.
Area of Science:
- Genetics and Genomics
- Molecular Biology
- Developmental Biology
Background:
- Transcription activator-like effector nucleases (TALENs) are advanced tools for targeted genome editing.
- Medaka fish (Oryzias latipes) are valuable vertebrate models for genetic and genomic research.
Purpose of the Study:
- To demonstrate efficient targeted mutagenesis in medaka using TALENs.
- To establish a method for rapidly generating knockout medaka strains.
- To investigate the DJ-1 gene and its protein function in medaka.
Main Methods:
- Design and construction of TALENs targeting the medaka DJ-1 gene.
- Injection of TALENs into medaka embryos to induce mutations.
- Analysis of mutations in injected embryos and subsequent generations (G0 and F1).
- Assessment of DJ-1 protein expression in mutated fish.
Main Results:
- TALENs induced targeted insertions and deletions (indels) in the medaka DJ-1 gene with high efficiency in a dose-dependent manner.
- TALEN-induced mutations were efficiently transmitted to the next generation (44-100% of G0 fish).
- Site-specific mutations were confirmed, with no detectable off-target mutations.
- DJ-1 protein was absent in medaka homozygous for TALEN-induced frameshift mutations.
- Specific N- and C-terminal regions of the TAL effector domain were identified as crucial for gene-disrupting activity.
Conclusions:
- TALEN technology provides an efficient and rapid method for generating knockout medaka strains.
- This study represents the first report of TALEN-mediated targeted mutagenesis in medaka.
- TALENs significantly enhance the utility of medaka as a model organism for genetic and genomic studies.
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