| RRC ID |
58479
|
| 著者 |
Huynh N, Wang S, King-Jones K.
|
| タイトル |
Spatial and temporal control of gene manipulation in Drosophila via drug-activated Cas9 nucleases.
|
| ジャーナル |
Insect Biochem Mol Biol
|
| Abstract |
Advances in CRISPR/Cas9 have revolutionized molecular biology and greatly facilitated the ability to manipulate gene function through the creation of precisely engineered mutants. We recently reported a collection of modular gateway-compatible Cas9/gRNA Drosophila lines to interfere with gene expression in a tissue-specific manner, including polytene tissues. However, most current in vivo CRISPR/Cas9 tools cannot temporally control the induction of Cas9 or gRNAs via external stimuli such as RU486. A drug-inducible CRISPR/Cas9 system would allow studying genes at later stages where early lethality is an issue. This would be especially useful when combined with tissue-specific expression of Cas9 or gRNAs, allowing for full spatiotemporal control. Here, we present a RU486-inducible version of Cas9 and also show that a Rapamycin-inducible Cas9, previously used in mammalian cell culture, works in Drosophila as well. Both RU486 and rapamycin-inducible Cas9 work in vivo and in Drosophila cell culture. We also present split Cas9 constructs for rapamycin-dependent gene disruption and activation. These approaches establish drug-inducible and thus temporally controlled CRISPR/Cas9 tools for gene disruption and expression in a living model organism. Our CRISPR/Cas9 vector collection can be easily adapted for any tissue and provides higher fidelity compared to RNAi approaches.
|
| 巻・号 |
120
|
| ページ |
103336
|
| 公開日 |
2020-5-1
|
| DOI |
10.1016/j.ibmb.2020.103336
|
| PII |
S0965-1748(20)30025-4
|
| PMID |
32105778
|
| MeSH |
Animals
Base Sequence
CRISPR-Cas Systems*
Drosophila / genetics*
Endonucleases
Gene Editing*
Gene Expression*
|
| IF |
3.827
|
| 引用数 |
0
|
| リソース情報 |
| ショウジョウバエ |
TBX-0004
TBX-0007
TBX-0008
TBX-0009 |
