Stringent analysis of gene function and protein-protein interactions using fluorescently tagged genes

Genetics. 2012 Mar;190(3):931-40. doi: 10.1534/genetics.111.136465. Epub 2011 Dec 14.


In Drosophila collections of green fluorescent protein (GFP) trap lines have been used to probe the endogenous expression patterns of trapped genes or the subcellular localization of their protein products. Here, we describe a method, based on nonoverlapping, highly specific, shRNA transgenes directed against GFP, that extends the utility of these collections to loss-of-function studies. Furthermore, we used a MiMIC transposon to generate GFP traps in Drosophila cell lines with distinct subcellular localization patterns, which will permit high-throughput screens using fluorescently tagged proteins. Finally, we show that fluorescent traps, paired with recombinant nanobodies and mass spectrometry, allow the study of endogenous protein complexes in Drosophila.

Publication types

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

MeSH terms

  • Animals
  • Cell Line
  • Cell Survival / genetics
  • Drosophila / genetics*
  • Drosophila / metabolism*
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism*
  • Embryo, Nonmammalian / metabolism
  • Female
  • Fluorescent Dyes*
  • Gene Order
  • Gene Silencing
  • Green Fluorescent Proteins* / genetics
  • Green Fluorescent Proteins* / metabolism
  • Multiprotein Complexes / isolation & purification
  • Multiprotein Complexes / metabolism
  • Peptide Elongation Factors / genetics
  • Peptide Elongation Factors / metabolism
  • Protein Binding / physiology
  • Protein Interaction Mapping / methods*
  • RNA, Small Interfering / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Stem Cells / metabolism


  • Drosophila Proteins
  • Fluorescent Dyes
  • Multiprotein Complexes
  • Peptide Elongation Factors
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Spt6 protein, Drosophila
  • Green Fluorescent Proteins