Bimolecular Fluorescence Complementation (BiFC) Analysis: Advances and Recent Applications for Genome-Wide Interaction Studies

J Mol Biol. 2015 Jun 5;427(11):2039-2055. doi: 10.1016/j.jmb.2015.03.005. Epub 2015 Mar 12.


Complex protein networks are involved in nearly all cellular processes. To uncover these vast networks of protein interactions, various high-throughput screening technologies have been developed. Over the last decade, bimolecular fluorescence complementation (BiFC) assay has been widely used to detect protein-protein interactions (PPIs) in living cells. This technique is based on the reconstitution of a fluorescent protein in vivo. Easy quantification of the BiFC signals allows effective cell-based high-throughput screenings for protein binding partners and drugs that modulate PPIs. Recently, with the development of large screening libraries, BiFC has been effectively applied for genome-wide PPI studies and has uncovered novel protein interactions, providing new insight into protein functions. In this review, we describe the development of reagents and methods used for BiFC-based screens in yeast, plants, and mammalian cells. We also discuss the advantages and drawbacks of these methods and highlight the application of BiFC in large-scale studies.

Keywords: bimolecular fluorescence complementation (BiFC) analysis; fluorescent proteins; genomics; proteomics.

Publication types

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

MeSH terms

  • Animals
  • Arabidopsis Proteins / metabolism
  • Fluorescent Dyes / metabolism
  • Genome-Wide Association Study
  • Green Fluorescent Proteins / metabolism
  • High-Throughput Screening Assays / methods*
  • Humans
  • Mammals
  • Molecular Probe Techniques*
  • Protein Interaction Mapping / methods*
  • Proteomics / methods
  • Telomere / metabolism
  • Yeasts / cytology
  • Yeasts / metabolism


  • Arabidopsis Proteins
  • Cyan Fluorescent Protein
  • Fluorescent Dyes
  • Green Fluorescent Proteins