An Efficient FLP-Based Toolkit for Spatiotemporal Control of Gene Expression in Caenorhabditis elegans

Genetics. 2017 Aug;206(4):1763-1778. doi: 10.1534/genetics.117.201012. Epub 2017 Jun 23.

Abstract

Site-specific recombinases are potent tools to regulate gene expression. In particular, the Cre (cyclization recombination) and FLP (flipase) enzymes are widely used to either activate or inactivate genes in a precise spatiotemporal manner. Both recombinases work efficiently in the popular model organism Caenorhabditis elegans, but their use in this nematode is still only sporadic. To increase the utility of the FLP system in C. elegans, we have generated a series of single-copy transgenic strains that stably express an optimized version of FLP in specific tissues or by heat induction. We show that recombination efficiencies reach 100% in several cell types, such as muscles, intestine, and serotonin-producing neurons. Moreover, we demonstrate that most promoters drive recombination exclusively in the expected tissues. As examples of the potentials of the FLP lines, we describe novel tools for induced cell ablation by expression of the PEEL-1 toxin and a versatile FLP-out cassette for generation of GFP-tagged conditional knockout alleles. Together with other recombinase-based reagents created by the C. elegans community, this toolkit increases the possibilities for detailed analyses of specific biological processes at developmental stages inside intact animals.

Keywords: DamID; FLP-out gene inactivation; MosSCI; PEEL-1 toxin; baf-1; genome engineering; tissue-specific gene expression.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans / growth & development
  • Caenorhabditis elegans / metabolism
  • DNA Nucleotidyltransferases / genetics
  • DNA Nucleotidyltransferases / metabolism
  • Gene Expression Regulation, Developmental*
  • Genes, Reporter
  • Genetic Engineering / methods*
  • Integrases / genetics
  • Integrases / metabolism
  • Intestinal Mucosa / metabolism
  • Muscle, Skeletal / metabolism
  • Neurons / metabolism
  • Recombination, Genetic
  • Transgenes

Substances

  • Bacterial Proteins
  • Cre recombinase
  • DNA Nucleotidyltransferases
  • FLP recombinase
  • Integrases