論文 - 詳細
| RRC ID | 56907 |
|---|---|
| 著者 | Takasu Y, Kobayashi I, Tamura T, Uchino K, Sezutsu H, Zurovec M. |
| タイトル | Precise genome editing in the silkworm Bombyx mori using TALENs and ds- and ssDNA donors - A practical approach. |
| ジャーナル | Insect Biochem Mol Biol |
| Abstract |
Engineered nucleases are able to introduce double stranded breaks at desired genomic locations. The breaks can be repaired by an error-prone non-homologous end joining (NHEJ) mechanism, or the repair process can be exploited to introduce precise DNA modifications by homology-directed repair (HDR) when provided with a suitable donor template. We designed a series of DNA donors including long dsDNA plasmids as well as short ssDNA oligonucleotides and compared the effectiveness of their utilization during gene targeting with highly efficient transcription activator-like effector nucleases (TALENs). While the use of long dsDNA donors for the incorporation of larger DNA fragments in Bombyx is still a problem, short single-stranded oligodeoxynucleotides (ssODNs) are incorporated quite efficiently. We show that appropriately designed ssODNs were integrated into germ cells in up to 79% of microinjected individuals and describe in more detail the conditions for the precise genome editing of Bombyx genes. We specify the donor sequence requirements that affected knock-in efficiency, and demonstrate the successful applications of this method of sequence deletion, insertion and replacement in the Bombyx genome. |
| 巻・号 | 78 |
| ページ | 29-38 |
| 公開日 | 2016-11-1 |
| DOI | 10.1016/j.ibmb.2016.08.006 |
| PII | S0965-1748(16)30124-2 |
| PMID | 27569417 |
| MeSH | Animals Bombyx / genetics* DNA / genetics* DNA / metabolism DNA, Single-Stranded / genetics* DNA, Single-Stranded / metabolism Gene Editing / methods* Transcription Activator-Like Effector Nucleases / genetics* Transcription Activator-Like Effector Nucleases / metabolism |
| IF | 3.618 |
| 引用数 | 9 |
| 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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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 3 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| カイコ | p50T w1-pnd |