Description
Product Usage Information
CST recommends transfection with 100 nM SignalSilence ® mTOR siRNA I 48 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use. Each vial contains the equivalent of 100 transfections, which corresponds to a final siRNA concentration of 100 nM per transfection in a 24-well plate with a total volume of 300 μl per well.
Storage
SignalSilence ® siRNA is supplied in RNAse-free water. Aliquot and store at -20ºC.
Background
The mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase (1-3) that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth (4,5). When sufficient nutrients are available, mTOR responds to a phosphatidic acid-mediated signal to transmit a positive signal to p70 S6 kinase and participate in the inactivation of the eIF4E inhibitor, 4E-BP1 (6). These events result in the translation of specific mRNA subpopulations. mTOR is phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway and autophosphorylated at Ser2481 (7,8). mTOR plays a key role in cell growth and homeostasis and may be abnormally regulated in tumors. For these reasons, mTOR is currently under investigation as a potential target for anti-cancer therapy (9). siRNA has been used to silence mTOR expression in 293-HEK cells causing decreased raptor expression, decreased p70 S6 kinase phosphorylation, and reduced cell size (9).
Alternate Names
FK506; FKBP-rapamycin; FRAP; FRAP1; RAFT1; rapamycin; RNAi; Signal Silence; siRNA; TOR; transfection
