A "FLP-Out" system for controlled gene expression in Caenorhabditis elegans.
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.
Voutev
Roumen
R
Department of Biology, Developmental Genetics Program, New York University, New York, New York 10003, USA.
Hubbard
E Jane Albert
EJ
D000483:Alleles; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005821:Genetic Techniques; D005822:Genetic Vectors; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D014157:Transcription Factors; D014158:Transcription, Genetic
D029742:Caenorhabditis elegans Proteins; D014157:Transcription Factors; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase