PUBLICATION
Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2
- Authors
- Pathak, G.P., Spiltoir, J.I., Höglund, C., Polstein, L.R., Heine-Koskinen, S., Gersbach, C.A., Rossi, J., Tucker, C.L.
- ID
- ZDB-PUB-171205-15
- Date
- 2017
- Source
- Nucleic acids research 45: e167 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Arabidopsis/genetics
- Arabidopsis Proteins/genetics*
- Basic Helix-Loop-Helix Transcription Factors/genetics*
- Cell Line
- Cryptochromes/genetics*
- DNA-Binding Proteins/genetics
- Gene Expression Regulation*
- HEK293 Cells
- Humans
- Light
- Optogenetics/methods*
- Transcriptional Activation/genetics*
- Zebrafish/genetics
- PubMed
- 28431041 Full text @ Nucleic Acids Res.
Citation
Pathak, G.P., Spiltoir, J.I., Höglund, C., Polstein, L.R., Heine-Koskinen, S., Gersbach, C.A., Rossi, J., Tucker, C.L. (2017) Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2. Nucleic acids research. 45:e167.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping