Originally published as Genetics Published Articles Ahead of Print on August 24, 2008.

Genetics, Vol. 180, 103-119, September 2008, Copyright © 2008
doi:10.1534/genetics.108.090274

A "FLP-Out" System for Controlled Gene Expression in Caenorhabditis elegans

Roumen Voutev^* and E. Jane Albert Hubbard^,1

^* Department of Biology, Developmental Genetics Program, New York University, New York, New York 10003 and Department of Pathology, Helen and Martin Kimmel Center for Stem Cell Biology and Developmental Genetics Program, Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, New York 10016

¹ Corresponding author: New York University School of Medicine, Skirball Institute of Biomolecular Medicine, 4th Floor, Lab 7, 540 First Ave., New York, NY 10016.
E-mail: jane.hubbard{at}med.nyu.edu

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.

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