Androgen signaling

Androgens, mainly testosterone and 5alpha-dihydrotestosterone (DHT) through their associations with Androgen receptor (AR, also known as NR3C4) will become the ctivated AR upon ligand binding undergoes conformational change to form a homodimer and interacts tightly with the Androgen Response Element (A-RE) which regulates gene expression.

Various regulators that regulate androgen induced apoptosis include BRCA1 and Smad3 and Akt, and much more.

 Androgen pathway detection

1. Reporting lentivirusses can monitor Androgen signaling pathway, the transcriptional activity of the androgen receptor.
2. The 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 Androgen Response Element (A-RE) sequence motif (5′- TGGAGGAACATATTGTATTTATT). The luciferase signal can be measured via Luciferase assay.
3. The fluorescent reporter can be detected via its fluorescent signal.
4. 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.
5. When androgenic hormones, testosterone or dihydrotestosterone binds to its receptor (AR), it activated the receptor.
6. The activated receptor then binds to A-RE which induce the downstream reporter’s expression as the result of activation for the minimal promoter. The reporter’s signal can be easily and rapidly readout via all types of assays (Reporter assay or by fluorescent microscope or FACS sorter).
7. Androgen Pathway Report Lentiviral particles in hek 293 cells.
8. 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.