Multi-chromatic control of mammalian gene expression and signaling

Nucleic Acids Res. 2013 Jul;41(12):e124. doi: 10.1093/nar/gkt340. Epub 2013 Apr 26.

Abstract

The emergence and future of mammalian synthetic biology depends on technologies for orchestrating and custom tailoring complementary gene expression and signaling processes in a predictable manner. Here, we demonstrate for the first time multi-chromatic expression control in mammalian cells by differentially inducing up to three genes in a single cell culture in response to light of different wavelengths. To this end, we developed an ultraviolet B (UVB)-inducible expression system by designing a UVB-responsive split transcription factor based on the Arabidopsis thaliana UVB receptor UVR8 and the WD40 domain of COP1. The system allowed high (up to 800-fold) UVB-induced gene expression in human, monkey, hamster and mouse cells. Based on a quantitative model, we determined critical system parameters. By combining this UVB-responsive system with blue and red light-inducible gene control technology, we demonstrate multi-chromatic multi-gene control by differentially expressing three genes in a single cell culture in mammalian cells, and we apply this system for the multi-chromatic control of angiogenic signaling processes. This portfolio of optogenetic tools enables the design and implementation of synthetic biological networks showing unmatched spatiotemporal precision for future research and biomedical applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arabidopsis Proteins / metabolism
  • CHO Cells
  • COS Cells
  • Cells, Cultured
  • Chlorocebus aethiops
  • Chromosomal Proteins, Non-Histone / metabolism
  • Cricetinae
  • Cricetulus
  • Gene Expression Regulation / radiation effects*
  • Humans
  • Mice
  • Neovascularization, Physiologic / genetics
  • Neovascularization, Physiologic / radiation effects
  • Signal Transduction / radiation effects*
  • Tissue Engineering
  • Ubiquitin-Protein Ligases / metabolism
  • Ultraviolet Rays*

Substances

  • Arabidopsis Proteins
  • Chromosomal Proteins, Non-Histone
  • Uvr8 protein, Arabidopsis
  • AT2G32950 protein, Arabidopsis
  • Ubiquitin-Protein Ligases