Target gene abundance contributes to the efficiency of siRNA-mediated gene silencing

Nucleic Acid Ther. 2014 Jun;24(3):192-8. doi: 10.1089/nat.2013.0466. Epub 2014 Feb 14.

Abstract

The gene-silencing activity of a small interfering RNA (siRNA) is determined by various factors. Considering that RNA interference (RNAi) is an unparalleled technology in both basic research and therapeutic applications, thorough understanding of the factors determining RNAi activity is critical. This report presents observations that siRNAs targeting KRT7 show cell-line-dependent activity, which correlates with the expression level of KRT7 mRNA. By modulating the target mRNA level, it was confirmed that highly expressed genes are more susceptible to siRNA-mediated gene silencing. Finally, several genes that show different expression levels in a cell-line dependent manner were tested, which verified the expression-level-dependent siRNA activities. These results strongly suggest that the abundance of target mRNA is a critical factor that determines the efficiency of the siRNA-mediated gene silencing in a given cellular context. This report should provide practical guidelines for designing RNAi experiments and for selecting targetable genes in RNAi therapeutics studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Gene Dosage*
  • Gene Expression Regulation
  • Gene Silencing*
  • Genes, Reporter
  • Humans
  • Keratin-7 / antagonists & inhibitors
  • Keratin-7 / genetics*
  • Keratin-7 / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism
  • Organ Specificity
  • RNA, Messenger / antagonists & inhibitors
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / genetics*
  • RNA, Small Interfering / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • KRT7 protein, human
  • Keratin-7
  • RNA, Messenger
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Luciferases