Journal
NATURE CHEMICAL BIOLOGY
Volume 10, Issue 3, Pages 196-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEMBIO.1430
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Funding
- US National Institutes of Health [R01 GM081875, GM106239, R01 GM103383, R01 GM096164]
- Cancer Prevention and Research Institute of Texas [RP130312]
- Defense Advanced Research Projects Agency (Living Foundries) [HR0011-12-C-0068]
- Robert A. Welch Foundation [I-1424]
- National Science Foundation
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Optogenetic gene expression systems can control transcription with spatial and temporal detail unequaled with traditional inducible promoter systems. However, current eukaryotic light-gated transcription systems are limited by toxicity, dynamic range or slow activation and deactivation. Here we present an optogenetic gene expression system that addresses these shortcomings and demonstrate its broad utility. Our approach uses an engineered version of EL222, a bacterial light-oxygen-voltage protein that binds DNA when illuminated with blue light. The system has a large (>100-fold) dynamic range of protein expression, rapid activation (<10 s) and deactivation kinetics (<50 s) and a highly linear response to light. With this system, we achieve light-gated transcription in several mammalian cell lines and intact zebrafish embryos with minimal basal gene activation and toxicity. Our approach provides a powerful new tool for optogenetic control of gene expression in space and time.
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