A "FLP-Out" system for controlled gene expression in Caenorhabditis elegans

Genetics. 2008 Sep;180(1):103-19. doi: 10.1534/genetics.108.090274. Epub 2008 Aug 24.

Abstract

We present a two-part system for conditional FLP-out of FRT-flanked sequences in Caenorhabditis elegans to control gene activity in a spatially and/or temporally regulated manner. Using reporters, we assess the system for efficacy and demonstrate its use as a cell lineage marking tool. In addition, we construct and test a dominant-negative form of hlh-12, a gene that encodes a basic helix-loop-helix (bHLH) transcription factor required for proper distal tip cell (DTC) migration. We show that this allele can be conditionally expressed from a heat-inducible FLP recombinase and can interfere with DTC migration. Using the same DTC assay, we conditionally express an hlh-12 RNAi-hairpin and induce the DTC migration defect. Finally, we introduce a set of traditional and Gateway-compatible vectors to facilitate construction of plasmids for this technology using any promoter, reporter, and gene/hairpin of interest.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Alleles
  • Animals
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans Proteins / genetics
  • DNA Nucleotidyltransferases / genetics*
  • Gene Expression Regulation
  • Genes, Reporter
  • Genetic Techniques*
  • Genetic Vectors
  • Models, Genetic
  • Plasmids / metabolism
  • Promoter Regions, Genetic
  • RNA Interference
  • Transcription Factors
  • Transcription, Genetic

Substances

  • Caenorhabditis elegans Proteins
  • Transcription Factors
  • DNA Nucleotidyltransferases
  • FLP recombinase