Abstract
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.
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Acknowledgments
We are grateful to J. Bonéy Montoya, A. Rao, D. Thorngren, and Y. Ziegler for assistance in the preparation of this manuscript. This work was supported by NIH grant R01 DK 53884 (to AMN).
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Curtis, C.D., Nardulli, A.M. (2009). Using RNA Interference to Study Protein Function. In: McEwan, I.J. (eds) The Nuclear Receptor Superfamily. Methods in Molecular Biology™, vol 505. Humana Press. https://doi.org/10.1007/978-1-60327-575-0_11
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DOI: https://doi.org/10.1007/978-1-60327-575-0_11
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