RNA干扰
基因沉默
基因表达
核糖核酸
小发夹RNA
基因
信使核糖核酸
遗传增强
病毒载体
表达式向量
载体(分子生物学)
作者
Eiji Morita,Jun Arii,Devin E. Christensen,Jörg Votteler,Wesley I. Sundquist
出处
期刊:BioTechniques
[Future Science Ltd]
日期:2012-08-01
卷期号:: 1-5
被引量:35
摘要
Transient transfection of small interfering RNA (siRNA) provides a powerful approach for studying cellular protein functions, particularly when the target protein can be re-expressed from an exogenous siRNA-resistant construct in order to rescue the knockdown phenotype, confirm siRNA target specificity, and support mutational analyses. Rescue experiments often fail, however, when siRNA-resistant constructs are expressed at suboptimal levels. Here, we describe an ensemble of mammalian protein expression vectors with CMV promoters of differing strengths. Using CHMP2A rescue of HIV-1 budding, we show that these vectors can combine high-transfection efficiencies with tunable protein expression levels to optimize the rescue of cellular phenotypes induced by siRNA transfection.
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