The B7-H4 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 B7-H4 (NM_ 024626.4) driven by a CMV promoter and a puromycin selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the B7-H4 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 #
Q7Z7D3
Background
B7-H4, also known as V-set domain-containing T-cell activation inhibitor 1 (VTCN1), is an immune checkpoint protein found on the surface of antigen-presenting cells. Its receptor has not yet been identified, but B7-H4 binds to receptor(s) on the surface of T cells, inhibiting T cell responses and suppressing tumor-associated immunity. High expression levels of B7-H4 have been correlated with tumor progression, particularly in ovarian and breast cancer.
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 a stable cell line expressing human B7-H4 with puromycin selection
Shipping Temperature
-80°C
Notes
To generate a B7-H4 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 antibiotic 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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