A system for stable expression of short interfering RNAs in mammalian cells

Science. 2002 Apr 19;296(5567):550-3. doi: 10.1126/science.1068999. Epub 2002 Mar 21.

Abstract

Mammalian genetic approaches to study gene function have been hampered by the lack of tools to generate stable loss-of-function phenotypes efficiently. We report here a new vector system, named pSUPER, which directs the synthesis of small interfering RNAs (siRNAs) in mammalian cells. We show that siRNA expression mediated by this vector causes efficient and specific down-regulation of gene expression, resulting in functional inactivation of the targeted genes. Stable expression of siRNAs using this vector mediates persistent suppression of gene expression, allowing the analysis of loss-of-function phenotypes that develop over longer periods of time. Therefore, the pSUPER vector constitutes a new and powerful system to analyze gene function in a variety of mammalian cell types.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaphase-Promoting Complex-Cyclosome
  • Cell Cycle Proteins
  • Down-Regulation
  • Gene Silencing*
  • Genes, p53
  • Genetic Techniques*
  • Genetic Vectors*
  • Humans
  • Ligases / genetics
  • Mutation
  • Nucleic Acid Conformation
  • Phenotype
  • Protein Kinases / genetics
  • Protein Kinases / metabolism
  • Protein-Serine-Threonine Kinases
  • Proto-Oncogene Proteins
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • RNA, Small Interfering
  • RNA, Untranslated / chemistry
  • RNA, Untranslated / genetics*
  • RNA, Untranslated / metabolism
  • Transfection
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 / metabolism
  • Ubiquitin-Protein Ligase Complexes*

Substances

  • Cell Cycle Proteins
  • Proto-Oncogene Proteins
  • RNA, Messenger
  • RNA, Small Interfering
  • RNA, Untranslated
  • Tumor Suppressor Protein p53
  • Ubiquitin-Protein Ligase Complexes
  • Anaphase-Promoting Complex-Cyclosome
  • Protein Kinases
  • Protein-Serine-Threonine Kinases
  • polo-like kinase 1
  • Ligases