MAPK/ERK Pathway

MAPK / ERK Signal Pathway: The mitogen-activated protein kinase (MAPK, also called ERK) pathway (also known as the Ras-Raf-MEK-ERK pathway) involves cell growth and differentiation and in the development of many cancers. Activated Ras activates the protein kinase activity of RAF kinase. RAF kinase phosphorylates and activates MEK (MEK1 and MEK2). MEK phosphorylates and activates MAPK. MAPK regulates the activities of several transcription factors. The MAPK phoshorylates Elk-1 protein which forms a complex with Serum Response Factor (SRF) and Ternary Complex Factor (TCR). This complex binds to Serum Response Element (SRE) sequence motif, causing transcriptional activation of target genes.

GenTarget developed a set of reporting lentivirus to monitor MAPK/ERK  signaling pathway. Those reporting lentivirus has a luminescent report (Luciferase, Renilla Luc) or a fluorescent report (GFP, RFP) or a  secreted SEAP report, under a minimal CMV promoter (mCMV)  that embedded with optimized tandem repeats of serum response element (SRE) sequence motif (5′- GGATGTCCATATTAGGA; CCAAAAAAGG). When transcription factors bind to SRE, the downstream reporter is expressed as the result of activation for the minimal promoter. This pathway lentivirus can be verified by FBS treatment.

The luciferase signal can be measured via Luciferase assay. The fluorescent reporter can be detected via its fluorescent signal. The SEAP secreted into cell culture supernatant, which allows to determine reporter activity without disturbing the cells, does not require the preparation of cell lysates and can be used for kinetic studies.

Those reporting lentivirus also contains a constitutively expressed fluorescent selection marker or an antibiotic selection marker under the RSV promoter, which makes it easier to select the stably infected signal reporting cells (to generate pathway specific sensor cell lines), or  provides internal reference for virus transduction efficiency when a fluorescent marker is under the RSV promoter. See lentivector’s core scheme below.

The premade, ready-to-use reporter lentivirus provides a much easier tool to monitor the activity of MAPK / ERK signaling pathways in virtually any mammalian cell type. It also allows to generate your own reporting cell line in your desired cell type for study or screen of pathway specific gene-knockdown, over-expression, or chemical / drug/protein treatment in the cell based assay.

Click the Product Manual (pdf) for all available products for this pathway reporter.