Title | Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2. |
Publication Type | Journal Article |
Year of Publication | 2017 |
Authors | GP Pathak, JI Spiltoir, C Höglund, LR Polstein, S Heine-Koskinen, CA Gersbach, J Rossi, and CL Tucker |
Journal | Nucleic acids research |
Volume | 45 |
Issue | 20 |
Start Page | e167 |
Date Published | 11/2017 |
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. |
DOI | 10.1093/nar/gkx260 |
Short Title | Nucleic acids research |