The GPRC5D 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 GPRC5D (NM_018654.1) driven by a CMV promoter and a Hygromycin selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the GPRC5D Lentivirus
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Synonyms
G-protein coupled receptor family C group 5 member D
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 #
Q9NZD1
Background
G protein-coupled receptor class C group 5 member D (GPRC5D) is an orphan receptor of little-known function belonging to the large family of G protein-coupled receptors. The protein is highly expressed in malignant plasma cells such multiple myeloma and is now considered a therapeutic target for antibody-based treatment of multiple myeloma. For example, CAR T cells and GPCR5DxCD3 bispecific antibodies are 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 GPRC5D with hygromycin selection
Shipping Temperature
-80°C
Notes
To generate a GPRC5D stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of hygromycin (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 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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