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
|
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) |