Effective RNA interference in cultured silkworm cells mediated by overexpression of Caenorhabditis elegans SID-1

RNA Biol. 2012 Jan;9(1):40-6. doi: 10.4161/rna.9.1.18084. Epub 2012 Jan 1.

Abstract

RNA interference (RNAi) is a conserved mechanism that catalyzes sequence-specific gene silencing and has been used for loss-of-function genetic screens in many organisms. Here, we demonstrated that the expression of Caenorhabditis elegans SID-1 (CeSID-1) could trigger effective gene silencing in the cultured silkworm cell line, BmN4 (BmN4-SID1). Soaking the BmN4-SID1 in dsRNA corresponding to endogenous target genes induced a significant decrease of the amount of mRNA or protein. A small amount of dsRNA was enough to silence the target gene in a few days. Overexpression of CeSID-1 did not affect the cell viability. Our results suggest that BmN4-SID1 can be used in many applications in silkworm cells and will become a valuable resource for gene analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bombyx / cytology
  • Bombyx / genetics*
  • Bombyx / metabolism
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism*
  • Cell Line
  • Cell Survival
  • Cloning, Molecular
  • Flow Cytometry
  • Gene Expression Regulation*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Plasmids / genetics
  • Plasmids / metabolism
  • Promoter Regions, Genetic
  • RNA Interference*
  • RNA, Double-Stranded / genetics
  • RNA, Double-Stranded / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Transfection

Substances

  • Caenorhabditis elegans Proteins
  • Membrane Proteins
  • RNA, Double-Stranded
  • RNA, Messenger
  • SID-1 protein, C elegans