The PSMA Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. The particles contain a human PSMA (NM_ 004476.3) driven by a CMV promoter and a puromycin selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the PSMA Lentivirus.
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.
Formulation
The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
UniProt #
Q04609
Background
PSMA (prostate-specific membrane antigen, also known as Folate hydrolase 1, FOLH1), is highly expressed in prostate cancer cells and is used as a diagnostic and prognostic indicator of prostate cancer. The enzyme has folate hydrolase and peptidase activity. It plays a role in prostate cancer progression through the PI3K-Akt and MAPK-ERK1/2 pathways. PSMA/FOLH1is used in the clinic as a target for PET (positron emission tomography) imaging of prostate cancer whereas radioactive analogs are used for the treatment of prostate cancer. It is also expressed in other tumor types and in a few normal tissues. PSMA/FOLH1 is the target of CAR-T cells and bi-specific antibodies currently under development.
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.
Applications
Generate stable cell line expressing human PSMA with puromycin selection
Shipping Temperature
-80°C
Notes
To generate a PSMA stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve) for antibiotics selection of transduced cells. Visit: https://bpsbioscience.com/cell-line-faq for guidelines on performing a kill curve.
Biosafety The 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.
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