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Intensive RNAi with lentiviral vectors in mammalian cells.

机译:在哺乳动物细胞中带有慢病毒载体的密集RNAi。

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摘要

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.
机译:RNAi是用于分析人类细胞中基因功能的强大技术。然而,靶基因mRNA的敲低不足或在没有严格控制的情况下影响的解释会损害其效用。我们回顾了基于慢病毒载体的方法,该方法能够进行瞬时或稳定的敲低以追踪人类CD4 + T细胞系和其他靶标中的mRNA水平。审查了重要的控制措施,包括通过重新表达靶基因来挽救预敲除前的表型。从考虑潜在的击倒靶点到分析表型的时间可能短短几个星期。

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