期刊
NUCLEIC ACIDS RESEARCH
卷 45, 期 20, 页码 -出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkx260
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资金
- National Science Foundation [CBET-1151035]
- National Institutes of Health [GM100225, DA036865]
- Academy of Finland [260030]
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1151035] Funding Source: National Science Foundation
- Academy of Finland (AKA) [260030, 260030] Funding Source: Academy of Finland (AKA)
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.
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