| RRC ID |
86764
|
| Author |
Pathak GP, Spiltoir JI, Höglund C, Polstein LR, Heine-Koskinen S, Gersbach CA, Rossi J, Tucker CL.
|
| Title |
Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2.
|
| Journal |
Nucleic Acids Res
|
| Abstract |
Optogenetic tools allow regulation of cellular processes with light, which can be delivered with spatiotemporal resolution. In previous work, we used cryptochrome 2 (CRY2) and CIB1, Arabidopsis proteins that interact upon light illumination, to regulate transcription with light in yeast. While adopting this approach to regulate transcription in mammalian cells, we observed light-dependent redistribution and clearing of CRY2-tethered proteins within the nucleus. The nuclear clearing phenotype was dependent on the presence of a dimerization domain contained within the CRY2-fused transcriptional activators. We used this knowledge to develop two different approaches to regulate cellular protein levels with light: a system using CRY2 and CIB1 to induce protein expression with light through stimulation of transcription, and a system using CRY2 and a LOV-fused degron to simultaneously block transcription and deplete protein levels with light. These tools will allow precise, bi-directional control of gene expression in a variety of cells and model systems.
|
| Volume |
45(20)
|
| Pages |
e167
|
| Published |
2017-11-16
|
| DOI |
10.1093/nar/gkx260
|
| PII |
3744534
|
| PMID |
28431041
|
| PMC |
PMC5714224
|
| MeSH |
Animals
Animals, Genetically Modified
Arabidopsis / genetics
Arabidopsis Proteins / genetics*
Basic Helix-Loop-Helix Proteins / genetics*
Cell Line
Cryptochromes / genetics*
DNA-Binding Proteins / genetics
Gene Expression Regulation*
HEK293 Cells
Humans
Light
Optogenetics / methods*
Transcriptional Activation / genetics*
Zebrafish / genetics
|
| IF |
11.502
|
| Resource |
| Zebrafish |
HGn39D |
