| RRC ID |
56342
|
| 著者 |
Kina H, Yoshitani T, Hanyu-Nakamura K, Nakamura A.
|
| タイトル |
Rapid and efficient generation of GFP-knocked-in Drosophila by the CRISPR-Cas9-mediated genome editing.
|
| ジャーナル |
Dev Growth Differ
|
| Abstract |
The CRISPR-Cas9 technology has been a powerful means to manipulate the genome in a wide range of organisms. A series of GFP knocked-in (GFPKI ) Drosophila strains have been generated through CRISPR-Cas9-induced double strand breaks coupled with homology-directed repairs in the presence of donor plasmids. They visualized specific cell types or intracellular structures in both fixed and live specimen. We provide a rapid and efficient strategy to identify KI lines. This method requires neither co-integration of a selection marker nor prior establishment of sgRNA-expressing transgenic lines. The injection of the mixture of a sgRNA/Cas9 expression plasmid and a donor plasmid into cleavage stage embryos efficiently generated multiple independent KI lines. A PCR-based selection allows to identify KI fly lines at the F1 generation (approximately 4 weeks after injection). These GFPKI strains have been deposited in the Kyoto Drosophila stock center, and made freely available to researchers at non-profit organizations. Thus, they will be useful resources for Drosophila research.
|
| 巻・号 |
61(4)
|
| ページ |
265-275
|
| 公開日 |
2019-5-1
|
| DOI |
10.1111/dgd.12607
|
| PMID |
31037730
|
| MeSH |
Animals
CRISPR-Cas Systems / genetics*
Drosophila / genetics*
Gene Editing / methods*
Gene Knock-In Techniques / methods*
Green Fluorescent Proteins / genetics*
Time Factors
|
| IF |
1.638
|
| 引用数 |
1
|
| リソース情報 |
| ショウジョウバエ |
DGRC#118616
DGRC#118617
DGRC#118618
DGRC#118619
DGRC#118620
DGRC#118621
DGRC#118622
DGRC#118623
DGRC#118624
DGRC#118625
DGRC#118626
DGRC#118651
DGRC#118652
DGRC#118653 |
