Journal
METHODS
Volume 47, Issue 4, Pages 298-303Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2008.11.001
Keywords
RNAi; Lentiviral vectors; ilvRNAi; shRNA; HIV; Knockdown
Funding
- NIAID NIH HHS [R01 AI077344, R01 AI077344-01A1, R01 AI077344-02] Funding Source: Medline
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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. (C) 2008 Elsevier Inc. All rights reserved.
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