E2F Pathway

The E2F signaling pathway plays a crucial role in cell cycle regulation and DNA synthesis in mammalian cells. E2F is a group of genes that encode a family of transcription factors, play a major role during the G1/S transition. E2F transcription factors can activate the transcription of genes involved in DNA replication and cell cycle progression  [Ref1].

2. Lentivirus Detects E2F Pathway:

GenTarget developed a set of reporting lentivirus to monitor E2F signaling pathway activity. Those reporting lentivirus has a luminescent report (Luciferase) or a fluorescent report (GFP), under a minimal CMV promoter (mCMV) that embedded with optimized tandem repeats of pathway specific E2F binding motif as the transcriptional response element (TRE ), sequence motif (5′- TTTCGCGC-TTTCGCGC-TTTCGCGC  3′) [Ref 2].

Upon the activation of E2F pathway (for example by adding specific growth factors, treating the cells with certain drugs), the E2F is activated and binds to E2F binding motif in the report’s promoter region, inducing the expression of the reporter gene, Luciferase or GFP which can then be easily measured. The amount of light produced (luminescence) or GFP fluorescent signal intensity, is proportional to the activity of the E2F pathway.

3. Structure of the Reporting Lentivirus:

The pathway specific reporting Lentivirus use either firefly-luciferase or GFP signal changes to detect the pathway’s activity.   Those reporting lentivirus also contains a constitutively expressed Puromycin antibiotic selection marker, or a GFP fluorescent selection marker, which makes it easier to select the stably transduced signal reporting cells, via Puromycin killing or GFP sorting respectively. See lentivector core scheme below.

The premade, ready-to-use reporter lentivirus provides a much easier tool to monitor the activity of the 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 screening 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.