Bidirectional approaches for optogenetic regulation of gene expression in mammalian cells using Arabidopsis cryptochrome 2
| Autor: | Camilla Höglund, Lauren R. Polstein, Chandra L. Tucker, Jessica I. Spiltoir, Jari Rossi, Sari Heine-Koskinen, Gopal P Pathak, Charles A. Gersbach |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Transcriptional Activation animal structures Light Arabidopsis DNA-binding protein Cell Line Animals Genetically Modified 03 medical and health sciences 0302 clinical medicine Cryptochrome Transcription (biology) Gene expression Genetics Transcriptional regulation Basic Helix-Loop-Helix Transcription Factors Animals Humans Zebrafish Regulation of gene expression biology Arabidopsis Proteins biology.organism_classification Cell biology Cryptochromes DNA-Binding Proteins Optogenetics 030104 developmental biology HEK293 Cells Gene Expression Regulation Methods Online sense organs Degron 030217 neurology & neurosurgery |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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