Using RNA interference to study protein function.
Using RNA interference to study protein function.
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DOI：
10.1007/978-1-60327-575-0_11
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发表时间：
2009
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影响因子：
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通讯作者：
Nardulli, Ann M
中科院分区：
文献类型：
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作者：
Curtis, Carol D;Nardulli, Ann M
RNA干扰在确定内源性表达蛋白在其正常细胞环境中的功能方面非常有用。在这一章中，我们描述了一种方法，使用小干扰RNA（siRNA）敲低mRNA和蛋白质表达在培养的细胞，使一个假定的调控蛋白对基因表达的影响可以划定。还包括使用真实的时间定量PCR和Western分析评估siRNA程序的有效性的方法。
RNA interference can be extremely useful in determining the function of an endogenously-expressed protein in its normal cellular environment. In this chapter, we describe a method that uses small interfering RNA (siRNA) to knock down mRNA and protein expression in cultured cells so that the effect of a putative regulatory protein on gene expression can be delineated. Methods of assessing the effectiveness of the siRNA procedure using real time quantitative PCR and Western analysis are also included.