Enhanced Neuronal RNAi in C. Elegans Using SID-1

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2010 Jun 1

PMID 20512143

Citations 232

Authors

Andrea Calixto

Dattananda Chelur

Irini Topalidou

Xiaoyin Chen

Martin Chalfie

Affiliations

Soon will be listed here.

Abstract

We expressed SID-1, a transmembrane protein from Caenorhabditis elegans that is required for systemic RNA interference (RNAi), in C. elegans neurons. This expression increased the response of neurons to double-stranded (ds)RNA delivered by feeding. Mutations in the lin-15b and lin-35 genes enhanced this effect. Worms expressing neuronal SID-1 showed RNAi phenotypes when fed with bacteria expressing dsRNA for known neuronal genes and for uncharacterized genes with no previously known neuronal phenotypes. Neuronal expression of sid-1 decreased nonneuronal RNAi, suggesting that neurons expressing transgenic sid-1(+) served as a sink for dsRNA. This effect, or a sid-1(-) background, can be used to uncover neuronal defects for lethal genes. Expression of sid-1(+) from cell-specific promoters in sid-1 mutants results in cell-specific feeding RNAi. We used these strains to identify a role for integrin signaling genes in mechanosensation.

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References

Schmitz C, Kinge P, Hutter H . Axon guidance genes identified in a large-scale RNAi screen using the RNAi-hypersensitive Caenorhabditis elegans strain nre-1(hd20) lin-15b(hd126). Proc Natl Acad Sci U S A. 2007; 104(3):834-9. PMC: 1783400. DOI: 10.1073/pnas.0510527104. View

Simmer F, Moorman C, van der Linden A, Kuijk E, van den Berghe P, Kamath R . Genome-wide RNAi of C. elegans using the hypersensitive rrf-3 strain reveals novel gene functions. PLoS Biol. 2003; 1(1):E12. PMC: 212692. DOI: 10.1371/journal.pbio.0000012. View

Wang D, Kennedy S, Conte Jr D, Kim J, Gabel H, Kamath R . Somatic misexpression of germline P granules and enhanced RNA interference in retinoblastoma pathway mutants. Nature. 2005; 436(7050):593-7. DOI: 10.1038/nature04010. View

Hardin J, King R, Thomas-Virnig C, Raich W . Zygotic loss of ZEN-4/MKLP1 results in disruption of epidermal morphogenesis in the C. elegans embryo. Dev Dyn. 2008; 237(3):830-6. DOI: 10.1002/dvdy.21455. View

Mello C, Kramer J, Stinchcomb D, Ambros V . Efficient gene transfer in C.elegans: extrachromosomal maintenance and integration of transforming sequences. EMBO J. 1991; 10(12):3959-70. PMC: 453137. DOI: 10.1002/j.1460-2075.1991.tb04966.x. View

Hodgkin J . Male Phenotypes and Mating Efficiency in CAENORHABDITIS ELEGANS. Genetics. 1983; 103(1):43-64. PMC: 1202023. DOI: 10.1093/genetics/103.1.43. View

Poyurovsky M, Jacq X, Ma C, Karni-Schmidt O, Parker P, Chalfie M . Nucleotide binding by the Mdm2 RING domain facilitates Arf-independent Mdm2 nucleolar localization. Mol Cell. 2003; 12(4):875-87. DOI: 10.1016/s1097-2765(03)00400-3. View

Lehner B, Calixto A, Crombie C, Tischler J, Fortunato A, Chalfie M . Loss of LIN-35, the Caenorhabditis elegans ortholog of the tumor suppressor p105Rb, results in enhanced RNA interference. Genome Biol. 2006; 7(1):R4. PMC: 1431716. DOI: 10.1186/gb-2006-7-1-r4. View

Winston W, Sutherlin M, Wright A, Feinberg E, Hunter C . Caenorhabditis elegans SID-2 is required for environmental RNA interference. Proc Natl Acad Sci U S A. 2007; 104(25):10565-70. PMC: 1965553. DOI: 10.1073/pnas.0611282104. View

10.

Emtage L, Gu G, Hartwieg E, Chalfie M . Extracellular proteins organize the mechanosensory channel complex in C. elegans touch receptor neurons. Neuron. 2004; 44(5):795-807. DOI: 10.1016/j.neuron.2004.11.010. View

11.

Simmer F, Tijsterman M, Parrish S, Koushika S, Nonet M, Fire A . Loss of the putative RNA-directed RNA polymerase RRF-3 makes C. elegans hypersensitive to RNAi. Curr Biol. 2002; 12(15):1317-9. DOI: 10.1016/s0960-9822(02)01041-2. View

12.

Sieburth D, Chng Q, Dybbs M, Tavazoie M, Kennedy S, Wang D . Systematic analysis of genes required for synapse structure and function. Nature. 2005; 436(7050):510-7. DOI: 10.1038/nature03809. View

13.

Maduro M, Pilgrim D . Identification and cloning of unc-119, a gene expressed in the Caenorhabditis elegans nervous system. Genetics. 1995; 141(3):977-88. PMC: 1206859. DOI: 10.1093/genetics/141.3.977. View

14.

Tijsterman M, May R, Simmer F, Okihara K, Plasterk R . Genes required for systemic RNA interference in Caenorhabditis elegans. Curr Biol. 2004; 14(2):111-6. DOI: 10.1016/j.cub.2003.12.029. View

15.

Timmons L, Fire A . Specific interference by ingested dsRNA. Nature. 1998; 395(6705):854. DOI: 10.1038/27579. View

16.

Fraser A, Kamath R, Zipperlen P, Martinez-Campos M, Sohrmann M, Ahringer J . Functional genomic analysis of C. elegans chromosome I by systematic RNA interference. Nature. 2000; 408(6810):325-30. DOI: 10.1038/35042517. View

17.

Tabara H, Grishok A, Mello C . RNAi in C. elegans: soaking in the genome sequence. Science. 1998; 282(5388):430-1. DOI: 10.1126/science.282.5388.430. View

18.

Feinberg E, Hunter C . Transport of dsRNA into cells by the transmembrane protein SID-1. Science. 2003; 301(5639):1545-7. DOI: 10.1126/science.1087117. View

19.

Kennedy S, Wang D, Ruvkun G . A conserved siRNA-degrading RNase negatively regulates RNA interference in C. elegans. Nature. 2004; 427(6975):645-9. DOI: 10.1038/nature02302. View

20.

Chalfie M, Au M . Genetic control of differentiation of the Caenorhabditis elegans touch receptor neurons. Science. 1989; 243(4894 Pt 1):1027-33. DOI: 10.1126/science.2646709. View