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Applications: Screen for activators or inhibitors of MAPK/ERK signaling pathway in transduced cells.Generate SRE TurboRFP reporter cell pools or stable cell lines following hygromycin selection.
Background: The MAPK (mitogen-activated protein kinase)/ERK (extracellular regulated kinase) signaling pathway is a major participant in the regulation of cell growth and differentiation. It can be activated by various extracellular stimuli including mitogens, growth factors, and cytokines. Upon stimulation, MEK1/2 (MAP kinase kinase 1/2) phosphorylate and activate ERK1/2. The activated ERK1/2 translocate to the nucleus where they phosphorylate and activate transcription factors. The TCFs (Ternary Complex Factors), including transcription factor Elk1 (ETS like-1 protein), are among the best-characterized substrates of ERK. When phosphorylated by ERK, Elk1 forms a complex with Serum Response Factor (SRF) and binds to the Serum Response Element (SRE), resulting in the expression of numerous mitogen-inducible genes. TurboRFP, a red (orange) fluorescent protein derived from the sea anemone Entacmaea quadricolor, has high photostability and pH stability allowing for easy assay readouts.
Description: SRE TurboRFP Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. The particles contain an TurboRFP (red fluorescent protein) reporter driven by the SRE (Serum Response Element) located upstream of the minimal TATA promoter. The lentiviruses also transduce a hygromycin selection gene (Figure 1). After transduction, activation of the MAPK (mitogen-activated protein kinase)/ERK (extracellular regulated kinase) signaling pathway in target cells can be monitored by assessing RFP expression.
Formulation: The lentiviruses were produced from HEK293T cells. Supplied in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations and produced at higher titers by special request, for an additional fee.
Storage Stability: Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.
Supplied As: Two vials (500 µl x 2) of SRE TurboRFP Reporter Lentivirus at ≥107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: BSL-2
References: Wong K.K., 2009 Recent Pat Anticancer Drug Discov. 4(1):28-35.
Treisman R., 1992 Trends Biochem Sci. 17(10): 423-426.
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