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Application: Screen and characterize modulators of the androgen receptor signaling pathway.Generate androgen receptor luciferase reporter cell pools or stable cell lines by puromycin selection.
Background: AR (androgen receptor), also known as NR3C4 (nuclear receptor family 3, group C, member 4) is a nuclear receptor involved in regulating gene expression. It is activated by testosterone and dihydrotestosterone, and it has been linked to prostate cancer. Prostate cancer is the most frequently diagnosed cancer and the second-leading cause of cancer death in American men. AR remains functional and is expressed in nearly all primary prostate cancers, with endocrine therapy aiming at reducing serum androgens and inhibiting AR. The androgen-induced transcriptional activation of AR is modulated by the interaction of AR with coregulators and by phosphorylation of AR and AR coregulators in response to growth factors. However, prostate cancer can almost always adapt to survive under castration levels of androgen or to AR inhibition. Castration resistance may involve AR point mutations, overexpression, changes in androgen biosynthesis, expression of constitutively active AR splice variants (active in the absence of ligand binding), and changes in androgen cofactors. As AR activity remains important in the progression of all stages of prostate cancer, AR continues to be an attractive molecular target of drugs against prostate cancer. 22Rv1 cell line is derived from a human prostate cancer xenograph propagated in mice and it is known to form tumors in nude mice. It is mostly dihydrotestosterone-independent for growth but responds to EGF (epidermal growth factor). They express androgen receptor (AR) that responds to androgen stimulation, making it one of the few available cellular models for prostate cancer studies.
Description: Androgen Receptor Luciferase Reporter Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce most mammalian cells, including primary and non-dividing cells. These viruses contain a firefly luciferase reporter driven by multiple copies of androgen response element (ARE) located upstream of the minimal TATA promoter. The lentiviruses also contain a puromycin selection marker (Figure 1). After transduction, the cellular androgen receptor signaling pathway can be monitored by measuring the luciferase activity.This product has been validated by testing the response to 5α-Dihydrotestosterone (5-DHT) of the transduced prostate cancer cell lines 22Rv1 and CWR-R1ca, but not LNcaP.
Formulation: The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations 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 lentivirus at a titer >107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Warnings: BiosafetyThe lentiviruses are produced with SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
Biosafety Level: BSL-2
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