A library of MiMICs allows tagging of genes and reversible, spatial and temporal knockdown of proteins in Drosophila

Elife. 2015 Mar 31:4:e05338. doi: 10.7554/eLife.05338.


Here, we document a collection of ∼7434 MiMIC (Minos Mediated Integration Cassette) insertions of which 2854 are inserted in coding introns. They allowed us to create a library of 400 GFP-tagged genes. We show that 72% of internally tagged proteins are functional, and that more than 90% can be imaged in unfixed tissues. Moreover, the tagged mRNAs can be knocked down by RNAi against GFP (iGFPi), and the tagged proteins can be efficiently knocked down by deGradFP technology. The phenotypes associated with RNA and protein knockdown typically correspond to severe loss of function or null mutant phenotypes. Finally, we demonstrate reversible, spatial, and temporal knockdown of tagged proteins in larvae and adult flies. This new strategy and collection of strains allows unprecedented in vivo manipulations in flies for many genes. These strategies will likely extend to vertebrates.

Keywords: D. melanogaster; RMCE; bruchpilot; cell biology; deGradFP; discs large 1; dunce; iGFPi; neuroscience.

Publication types

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

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Blotting, Western
  • Brain / metabolism
  • DNA Transposable Elements / genetics*
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / genetics*
  • Drosophila melanogaster / metabolism
  • Drosophila melanogaster / physiology
  • Gene Expression
  • Gene Library*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Larva / genetics
  • Larva / metabolism
  • Learning / physiology
  • Microscopy, Confocal
  • Mutagenesis, Insertional*
  • RNA Interference*
  • Time Factors
  • Tumor Suppressor Proteins / genetics
  • Tumor Suppressor Proteins / metabolism
  • alpha Catenin / genetics
  • alpha Catenin / metabolism


  • BRP protein, Drosophila
  • DNA Transposable Elements
  • Drosophila Proteins
  • Tumor Suppressor Proteins
  • alpha Catenin
  • dlg1 protein, Drosophila
  • Green Fluorescent Proteins