About JAK-STAT Signal Pathway:
The JAK-STAT signaling pathway transmits information from extracellular chemical signals to the nucleus resulting in target gene expression. INFa and INFß (interferon alpha, beta) is a Type I interferon. The extracellular signal (interferon) bind to their cell surface receptor, which leads to activation of Janus Kinase (JAK) which in turn activate two Signal Transducer and Activator of Transcription (STAT1/STAT2). The activated STAT1 and STAT2 form a protein complex with IRF9 / P48 (ISGF3) that binds to a general motif sequence, ISRE (Interferon Stimulated Response Element) in the promoter region of the interferon-inducible genes, which promotes those gene’s transcription. This JAK-STAT signal transduction pathways plays a critical role in cell growth, hematopoiesis, and immune response.
Detect JAK-STAT Signal Pathway:
GenTarget developed a set of reporting lentivirus to monitor interferon-induced 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 ISRE sequence motif (5′- TAGTTTCACTTTCCC). When induced transcription factors bind to ISRE sequence, the downstream reporter is expressed as the result of activation for the minimal promoter. This pathway lentivirus can be verified by Interferon Alpha (INFa) protein.
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.
Use JAK-STAT Signal Pathway Report Lentivirus:
Those reporting lentivirus 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 core scheme below.
The premade, ready-to-use reporter lentivirus provides a much easier tool to monitor the activity of JAK/STAT 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 (such as INFa protein) in the cell based assay.
Click the Product Manual (pdf) for all available products for this pathway reporter.