Conditional gene expression and RNAi using MEC-8-dependent splicing in C. elegans.
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.
Calixto
Andrea
A
Department of Biological Sciences, Columbia University, New York, New York, USA.
Ma
Charles
C
Chalfie
Martin
M
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D017238:Genes, Helminth; D007438:Introns / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics; D034622:RNA Interference / Q000502:physiology*; D012326:RNA Splicing / Q000502:physiology*; D016601:RNA-Binding Proteins / Q000502:physiology*; D013696:Temperature
D029742:Caenorhabditis elegans Proteins; C096705:MEC-2 protein, C elegans; D008565:Membrane Proteins; D016601:RNA-Binding Proteins; C099356:mec-8 protein, C elegans