Spatiotemporal control of gene expression in mammalian cells and in mice using the LightOn system

Curr Protoc Chem Biol. 2013;5(2):111-29. doi: 10.1002/9780470559277.ch120267.

Abstract

A light-switchable transgene system could be a powerful optogenetic tool for the precise manipulation of spatiotemporal gene expression in multicellular organisms. We have developed the LightOn system, which consists of a single chimeric protein (GAVPO) that can homodimerize and bind to promoters upon exposure to blue light, activating transcription of a target gene. This article describes protocols for precise control of gene expression in mammalian cells and mice using the LightOn system. These protocols can be carried out in an ordinary laboratory, as both liposome-mediated transfection and hydrodynamic tail vein injection are routine methods that can easily transfer the LightOn system to mammalian cells and mouse liver, respectively. The illumination equipment can also be easily obtained. The LightOn system can provide a robust, convenient means to control the expression of a gene of interest, with unprecedented temporal and spatial accuracy in manipulating an extremely broad range of biological processes.

Keywords: LightOn system; gene expression; light induced; mammalian cell; mice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Glucose / metabolism
  • Cell Line
  • Diabetes Mellitus, Experimental / blood
  • Diabetes Mellitus, Experimental / genetics
  • Gene Expression Regulation*
  • Genes, Reporter
  • Insulin / metabolism
  • Light*
  • Mice
  • Mice, Transgenic
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Tail / blood supply
  • Transfection / methods

Substances

  • Blood Glucose
  • Insulin
  • Recombinant Fusion Proteins