Reference - Detail
| RRC ID | 58479 |
|---|---|
| Author | Huynh N, Wang S, King-Jones K. |
| Title | Spatial and temporal control of gene manipulation in Drosophila via drug-activated Cas9 nucleases. |
| Journal | 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. |
| Volume | 120 |
| Pages | 103336 |
| Published | 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 |
| Times Cited | 0 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 3 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Drosophila | TBX-0004 TBX-0007 TBX-0008 TBX-0009 |