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.
Fraser
Iain
I
AfCS Molecular Biology Laboratory, Division of Biology, California Institute of Technology, Pasadena, CA, USA.
Liu
Wei
W
Rebres
Robert
R
Roach
Tamara
T
Zavzavadjian
Joelle
J
Santat
Leah
L
Liu
Jamie
J
Wall
Estelle
E
Mumby
Marc
M
D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008969:Molecular Sequence Data; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics
D034741:RNA, Small Interfering; D010749:Phosphoprotein Phosphatases