Ratiometric Biosensors Based on Dimerization-Dependent Fluorescent Protein Exchange

Nat Methods. 2015 Mar;12(3):195-8. doi: 10.1038/nmeth.3261. Epub 2015 Jan 26.

Abstract

We have developed a versatile new class of genetically encoded fluorescent biosensor based on reversible exchange of the heterodimeric partners of green and red dimerization-dependent fluorescent proteins. We demonstrate the use of this strategy to construct both intermolecular and intramolecular ratiometric biosensors for qualitative imaging of caspase activity, Ca(2+) concentration dynamics and other second-messenger signaling activities.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Biosensing Techniques / methods*
  • Calcium / metabolism
  • Calmodulin / metabolism
  • Caspase 3 / genetics
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Molecular Imaging / methods
  • Protein Multimerization

Substances

  • Calmodulin
  • Luminescent Proteins
  • red fluorescent protein
  • Green Fluorescent Proteins
  • Caspase 3
  • Calcium