Genetics. Published Articles Ahead of Print: August 24, 2008, Copyright © 2008
doi:10.1534/genetics.108.090274


A more recent version of this article appeared on September 1, 2008.

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A "FLP-out" system for controlled gene expression in C. elegans

Roumen Voutev 1 and E. Jane Albert Hubbard 2*

1 New York University
2 New York University School of Medicine

* To whom correspondence should be addressed. E-mail: jhubbard{at}saturn.med.nyu.edu.

Submitted on April 14, 2008
Revised on May 6, 2008
Accepted on 26 June 2008


Abstract

We present a two-part system for conditional FLP-out of FRT-flanked sequences in C. 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 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.

Key Words: FLP recombinase, FLP/FRT, RNAi, hlh-12, lineage