Conditional gene expression and RNAi using MEC-8-dependent splicing in C. elegans

Nat Methods. 2010 May;7(5):407-11. doi: 10.1038/nmeth.1445. Epub 2010 Apr 4.

Abstract

We describe a method for conditional regulation of gene expression based on the processing of an intron cassette. The RNA processing factor MEC-8 is necessary for the function of the Caenorhabditis elegans touch receptor neurons; mec-8 mutants are touch insensitive. We show here that this insensitivity involves the loss of MEC-8-dependent splicing of mec-2, which encodes a component of the mechanosensory transduction complex. MEC-8 is needed to remove the ninth intron in mec-2 pre-mRNA to form the longest of three mRNAs, mec-2a. Without MEC-8, splicing causes the termination of the transcript. Inclusion of mec-2 intron 9 is sufficient to convey mec-8-dependent regulation on other genes and, in mec-8(u218ts) mutants, resulted in their temperature-dependent expression. Because mec-8 is expressed ubiquitously in embryos and extensively in larvae, this system should produce temperature-sensitive expression for most genes. As an example, we report a strain that exhibits temperature-dependent RNA interference.

Publication types

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

MeSH terms

  • Animals
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / physiology*
  • Gene Expression Regulation, Developmental / physiology*
  • Genes, Helminth
  • Introns / physiology
  • Membrane Proteins / genetics
  • RNA Interference / physiology*
  • RNA Splicing / physiology*
  • RNA-Binding Proteins / physiology*
  • Temperature

Substances

  • Caenorhabditis elegans Proteins
  • MEC-2 protein, C elegans
  • Membrane Proteins
  • RNA-Binding Proteins
  • mec-8 protein, C elegans