The GPC3 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 GPC3 (Glypican 3, NM_ 004484.2) driven by an EF1A promoter and a puromycin selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the GPC3 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 #
P51654
Background
GPC3 (Glypican-3, also known as OCI5) is a cell surface proteoglycan highly expressed in the lungs, liver, and kidneys. Its function is tissue-dependent and can either promote or suppress tumorigenesis. Being a heparin sulfate proteoglycan, it is overexpressed in neoplasms including malignant melanoma, hepatocellular carcinoma, and testicular yolk sac tumors in which it regulates cell growth and differentiation. It is also involved in the negative regulation of the hedgehog signaling pathway and in the positive regulation of the non-canonical Wnt signaling pathway.
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 GPC3 with puromycin selection
Shipping Temperature
-80°C
Notes
To generate a GPC3 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 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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