Intensive RNAi with lentiviral vectors in mammalian cells

Tanner Miest, Dyana Saenz, Anne Meehan, Manuel Llano, Eric M. Poeschla

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

RNAi is a powerful technology for analyzing gene function in human cells. However, its utility can be compromised by inadequate knockdown of the target mRNA or by interpretation of effects without rigorous controls. We review lentiviral vector-based methods that enable transient or stable knockdowns to trace mRNA levels in human CD4+ T cell lines and other targets. Critical controls are reviewed, including rescue of the pre-knockdown phenotype by re-expression of the targeted gene. The time from thinking about a potential knockdown target to analysis of phenotypes can be as short as a few weeks.

Original languageEnglish (US)
Pages (from-to)298-303
Number of pages6
JournalMethods
Volume47
Issue number4
DOIs
StatePublished - Apr 2009

Keywords

  • HIV
  • Knockdown
  • Lentiviral vectors
  • RNAi
  • ilvRNAi
  • shRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)

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    Miest, T., Saenz, D., Meehan, A., Llano, M., & Poeschla, E. M. (2009). Intensive RNAi with lentiviral vectors in mammalian cells. Methods, 47(4), 298-303. https://doi.org/10.1016/j.ymeth.2008.11.001