論文 - 詳細
| RRC ID | 47703 |
|---|---|
| 著者 | Ansai S, Inohaya K, Yoshiura Y, Schartl M, Uemura N, Takahashi R, Kinoshita M. |
| タイトル | Design, evaluation, and screening methods for efficient targeted mutagenesis with transcription activator-like effector nucleases in medaka. |
| ジャーナル | Dev Growth Differ |
| Abstract |
Genome editing using engineered nucleases such as transcription activator-like effector nucleases (TALENs) has become a powerful technology for reverse genetics. In this study, we have described efficient detection methods for TALEN-induced mutations at endogenous loci and presented guidelines of TALEN design for efficient targeted mutagenesis in medaka, Oryzias latipes. We performed a heteroduplex mobility assay (HMA) using an automated microchip electrophoresis system, which is a simple and high-throughput method for evaluation of in vivo activity of TALENs and for genotyping mutant fish of F1 or later generations. We found that a specific pattern of mutations is dominant for TALENs harboring several base pairs of homologous sequences in target sequence. Furthermore, we found that a 5' T, upstream of each TALEN-binding sequence, is not essential for genomic DNA cleavage. Our findings provide information that expands the potential of TALENs and other engineered nucleases as tools for targeted genome editing in a wide range of organisms, including medaka. |
| 巻・号 | 56(1) |
| ページ | 98-107 |
| 公開日 | 2014-1-1 |
| DOI | 10.1111/dgd.12104 |
| PMID | 24286287 |
| MeSH | Animals Endodeoxyribonucleases / genetics Endodeoxyribonucleases / metabolism* Gene Targeting / methods* Heteroduplex Analysis Mutagenesis, Site-Directed / methods* Oryzias / genetics* |
| IF | 1.723 |
| 引用数 | 41 |
| WOS 分野 | DEVELOPMENTAL BIOLOGY CELL BIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| メダカ | d-rR/TOKYO (MT837) OK-Cab (MT830) DJ-1(delta7) (MT972) MC4R(delta7) (MT973) MC4R(delta11) (MT974) Sakyo (WS1164) |