| RRC ID |
29682
|
| 著者 |
Takasu Y, Sajwan S, Daimon T, Osanai-Futahashi M, Uchino K, Sezutsu H, Tamura T, Zurovec M.
|
| タイトル |
Efficient TALEN construction for Bombyx mori gene targeting.
|
| ジャーナル |
PLoS One
|
| Abstract |
Engineered nucleases are artificial enzymes able to introduce double stranded breaks at desired genomic locations. The double stranded breaks start the error-prone repair process of non-homologous end-joining (NHEJ), which eventually leads to the induction of mutations at target sites. We showed earlier that ZFNs and TALENs are able to induce NHEJ mutations in the B. mori genome. In order to optimize our mutagenesis protocol, we modified one of the reported truncated TALEN scaffolds and optimized it for use in the B. mori embryo. We also established a novel B. mori somatic cell assay suitable for the preselection of highly efficient TALENs directly in the B. mori model system. We compared the efficiency of several TALEN pairs based on three different frameworks using the BmBLOS2 gene. The new active TALENs show one order of magnitude higher efficiency than those we used previously. We confirmed the utility of our improved protocol by mutagenesis of the autosomal gene, red egg (Bm-re) and showed that it allows obtaining homozygous mutants in G1. Our procedure minimizes the chance of failure in B. mori gene targeting experiments.
|
| 巻・号 |
8(9)
|
| ページ |
e73458
|
| 公開日 |
2013-1-1
|
| DOI |
10.1371/journal.pone.0073458
|
| PII |
PONE-D-13-12139
|
| PMID |
24058473
|
| PMC |
PMC3776831
|
| MeSH |
Animals
Base Sequence
Bombyx / embryology
Bombyx / genetics*
DNA End-Joining Repair
Embryo, Nonmammalian
Endonucleases / genetics*
Gene Targeting / methods*
Genetic Engineering / methods*
Genome, Insect*
Homozygote
Larva / genetics
Larva / growth & development
Molecular Sequence Data
Mutagenesis, Insertional
Plant Proteins / genetics*
Transcription, Genetic
|
| IF |
2.74
|
| 引用数 |
44
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| カイコ |
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