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Nat Chem Biol. 2014 Mar;10(3):196-202. doi: 10.1038/nchembio.1430. Epub 2014 Jan 12.

An optogenetic gene expression system with rapid activation and deactivation kinetics.

Author information

1
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
2
1] Cardiovascular Research Institute, University of California-San Francisco, San Francisco, California, USA. [2] Department of Biochemistry and Biophysics, University of California-San Francisco, San Francisco, California, USA.
3
Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
4
Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
5
1] Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [2] Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA. [3] Structural Biology Initiative, Advanced Science Research Center, City University of New York, New York, New York, USA.

Abstract

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.

PMID:
24413462
PMCID:
PMC3944926
DOI:
10.1038/nchembio.1430
[Indexed for MEDLINE]
Free PMC Article

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