| 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
|
| リソース情報 |
| カイコ |
p50T
w1-pnd |
