SignalSilence® APC6 siRNA II from Cell Signaling Technology (CST) allows the researcher to specifically inhibit APC6 expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.
Quality Control
Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.
irections for Use
CST recommends transfection with 100 nM SignalSilence® APC6 siRNA II 48 to 72 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.
Background
The Adenomatous Polyposis Coli (APC) tumor suppressor gene is mutated in most familial and sporadic colorectal cancers and encodes a large cytoplasmic protein that is implicated in cell migration, cell adhesion, and proliferation (1). APC binds directly to microtubules and lack of APC leads to defective mitotic spindles and aneuploidy due to missegregation of chromosomes (2). APC is well characterized as a scaffolding protein, binds to β-catenin, and is involved in the regulation of its intracellular concentration. In the absence of a Wnt signal, GSK-3β phosphorylates all three members of the APC-β-catenin-axin complex and this phosphorylation of β-catenin creates a recognition site for ubiquitin, the signal for proteasome-mediated degradation. In the presence of a Wnt signal, dishevelled inactivates GSK-3β and β-catenin coordinates gene transcription of proteins important for the control of cell cycle progression and proliferation, such as cyclin D1 and c-Myc (3).