Optimized light-inducible transcription in mammalian cells using Flavin Kelch-repeat F-box1/GIGANTEA and CRY2/CIB1

Nucleic Acids Res. 2017 Nov 16;45(20):e172. doi: 10.1093/nar/gkx804.

Abstract

Light-inducible systems allow spatiotemporal control of a variety of biological activities. Here, we report newly optimized optogenetic tools to induce transcription with light in mammalian cells, using the Arabidopsis photoreceptor Flavin Kelch-repeat F-box 1 (FKF1) and its binding partner GIGANTEA (GI) as well as CRY2/CIB1. By combining the mutagenesis of FKF1 with the optimization of a split FKF1/GI dimerized Gal4-VP16 transcriptional system, we identified constructs enabling significantly improved light-triggered transcriptional induction. In addition, we have improved the CRY2/CIB1-based light-inducible transcription with split construct optimization. The improvements regarding the FKF1/GI- and CRY2/CIB1-based systems will be widely applicable for the light-dependent control of transcription in mammalian cells.

MeSH terms

  • 3T3 Cells
  • Animals
  • Arabidopsis / enzymology
  • Arabidopsis / genetics
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism
  • Basic Helix-Loop-Helix Transcription Factors / genetics*
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cell Line
  • Cryptochromes / genetics*
  • Cryptochromes / metabolism
  • Female
  • Gene Expression Regulation / genetics*
  • HEK293 Cells
  • Humans
  • Light
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Optogenetics / methods*
  • Protein Binding / genetics
  • Transcriptional Activation / genetics*

Substances

  • Arabidopsis Proteins
  • Basic Helix-Loop-Helix Transcription Factors
  • CIB1 protein, Arabidopsis
  • CRY2 protein, Arabidopsis
  • Cryptochromes
  • FKF1 protein, Arabidopsis
  • GI protein, Arabidopsis