Optogenetic control with a photocleavable protein, PhoCl

Nat Methods. 2017 Apr;14(4):391-394. doi: 10.1038/nmeth.4222. Epub 2017 Mar 13.

Abstract

To expand the range of experiments that are accessible with optogenetics, we developed a photocleavable protein (PhoCl) that spontaneously dissociates into two fragments after violet-light-induced cleavage of a specific bond in the protein backbone. We demonstrated that PhoCl can be used to engineer light-activatable Cre recombinase, Gal4 transcription factor, and a viral protease that in turn was used to activate opening of the large-pore ion channel Pannexin-1.

MeSH terms

  • Connexins / genetics
  • Connexins / metabolism
  • Directed Molecular Evolution
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • HSP90 Heat-Shock Proteins / genetics
  • HSP90 Heat-Shock Proteins / metabolism
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Nuclear Localization Signals / genetics
  • Optogenetics / methods*
  • Patch-Clamp Techniques
  • Photochemistry / methods
  • Protein Engineering / methods*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Red Fluorescent Protein
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / metabolism

Substances

  • Connexins
  • HSP90 Heat-Shock Proteins
  • Luminescent Proteins
  • NS3 protein, hepatitis C virus
  • Nerve Tissue Proteins
  • Nuclear Localization Signals
  • PANX1 protein, human
  • Recombinant Proteins
  • Viral Nonstructural Proteins
  • Green Fluorescent Proteins