A gene-specific T2A-GAL4 library for Drosophila

Elife. 2018 Mar 22;7:e35574. doi: 10.7554/eLife.35574.

Abstract

We generated a library of ~1000 Drosophila stocks in which we inserted a construct in the intron of genes allowing expression of GAL4 under control of endogenous promoters while arresting transcription with a polyadenylation signal 3' of the GAL4. This allows numerous applications. First, ~90% of insertions in essential genes cause a severe loss-of-function phenotype, an effective way to mutagenize genes. Interestingly, 12/14 chromosomes engineered through CRISPR do not carry second-site lethal mutations. Second, 26/36 (70%) of lethal insertions tested are rescued with a single UAS-cDNA construct. Third, loss-of-function phenotypes associated with many GAL4 insertions can be reverted by excision with UAS-flippase. Fourth, GAL4 driven UAS-GFP/RFP reports tissue and cell-type specificity of gene expression with high sensitivity. We report the expression of hundreds of genes not previously reported. Finally, inserted cassettes can be replaced with GFP or any DNA. These stocks comprise a powerful resource for assessing gene function.

Keywords: CRIMIC; D. melanogaster; GAL4/UAS; MiMIC; cassette excision; chromosomes; complementation; gene expression; loss of function.

Publication types

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

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / cytology
  • Drosophila melanogaster / genetics*
  • Drosophila melanogaster / metabolism
  • Gene Expression Profiling
  • Gene Library*
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mutagenesis, Insertional
  • Organ Specificity / genetics
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism

Substances

  • Drosophila Proteins
  • GAL4 protein, Drosophila
  • Luminescent Proteins
  • Transcription Factors