An optogenetic gene expression system with rapid activation and deactivation kinetics

Laura B. Motta-Mena, Anna Reade, Michael J. Mallory, Spencer Glantz, Orion D. Weiner, Kristen W. Lynch, Kevin H. Gardner

Research output: Contribution to journalArticlepeer-review

168 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)196-202
Number of pages7
JournalNature chemical biology
Volume10
Issue number3
DOIs
StatePublished - Mar 2014

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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