WormBase Tree Display for RNAi: WBRNAi00102072

WBRNAi00102072 Homol: Homol_homol: C54D10:RNAi
  Sequence_info: PCR_product: sjj_C54D10.8
  Experiment: Laboratory: JF
    Date: 13 Sep 2013 00:00:00
    Strain: WBStrain00028995
    Treatment: A two-generation approach was used to ensure gene knockdown throughout all C. elegans developmental stages. First, dsRNA-expressing bacterial cultures were grown overnight at 37 degrees Celsius with constant agitation. Isopropyl Beta-D-1-thiogalactopyranoside was added to a final concentration of 2 millimolar, and the incubation continued for 1 hour. Bacteria were then collected and resuspended in complete K-medium. Bacteria were added to appropriate wells in a 96-well plate, then nine L4 nematodes were added to each well, and incubated at 20 degrees Celsius for 48 hours. Following this incubation, 50 L1 larvae were transferred from each well to new 96-well plates, containing fresh dsRNA-expressing bacteria and mercuric chloride (HgCl2) or methylmercury chloride (MeHgCl). Nematodes were exposed to mercurial alone, gene-specific dsRNA alone, or mercurial and gene-specific dsRNA. The effects of dsRNA and/or mercurial on C. elegans growth were assessed following a 48 hour incubation. The initial assessment of gene-mercurial interactions was performed by visual observation. Any gene whose knockdown appeared to affect C. elegans growth, and thus a potential gene-mercurial interaction, was selected for additional analysis. All of the selected clones were sequenced to verify their identity. Of the 155 clones identified in the initial assessment, six were a different gene than described. In the second phase of the screen, nematodes were fed dsRNA-expressing bacteria as described above. Growth was then measured using the C. elegans growth assay, as previously described. A 2-way ANOVA was used to test for significant gene-mercury interactions using 500-800 nematodes per treatment condition. The criterion for a statistically significant interaction was p < 0.01.
    Temperature: 20
    Delivered_by: Bacterial_feeding
  Inhibits: Predicted_gene: C54D10.8b Inferred_automatically: RNAi_primary
      C54D10.8a Inferred_automatically: RNAi_primary
    Gene: WBGene00008302 Inferred_automatically: RNAi_primary
    Transcript: C54D10.8a.1 Inferred_automatically: RNAi_primary
      C54D10.8b.1 Inferred_automatically: RNAi_primary
  Species: Caenorhabditis elegans
  Reference: WBPaper00044316
  Phenotype: WBPhenotype:0001502 Remark: RNAi resulted in animals that exhibited increased resistance to exposure to mercuric chloride (HgCl2), compared to wild type controls
      Affected_by: Molecule: WBMol:00002810
  Remark: (Table 6, S7) C54D10.8 RNAi. Authors state RNAi clones obtained from the Ahringer and Vidal laboratories, Ahringer clone used for curation. If Ahringer clone not available, Vidal laboratory clone used for curation.
  Method: RNAi