The Use of RNA Interference to Analyze Protein Phosphatase Function in Mammalian Cells
The use of RNA interference to knock down protein phosphatases has proven to be a valuable approach to understanding the functions of these enzymes in mammalian cells. Many protein phosphatases exist as multisubunit and multigene families, which has made it difficult to assess their physiological functions using traditional approaches. The ability to selectively knock down specific subunits and individual isoforms with RNA interference has begun to make it possible to determine the contributions of individual phosphatase proteins to cellular signaling. This chapter describes methods for knocking down protein phosphatases with small interfering RNAs in easily transfectable cells and by the introduction of short-hairpin RNAs into less tractable cells using lentivirus vectors.
Key WordsProtein serine/threonine phosphatase protein tyrosine phosphatase protein phosphatase 2A PP2A SH2-containing protein tyrosine phosphatase 2 Shp2 RNA interference small interfering RNA short hairpin RNA lentivirus transfection transduction physiological function
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