The Nectin-4 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 Nectin-4 (NM_ 030916.2) driven by an EF1A promoter and a puromycin selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the Nectin-4 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 #
Q96NY8
Background
In contrast to other Nectins, which are found extensively in adult tissues, Nectin4 is abundant during fetal development but declines in adult life. However, its expression is elevated specifically in lung, breast, pancreas, and ovarian cancers, and is associated with poor prognosis. Nectin4 modulates the expression of proteins involved in the epithelial–mesenchymal transition via the PI3K/AKT pathway. In addition, Nectin4 ligand for TIGIT in cancer cells. It is considered a promising target for cancer immunotherapy.
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 Nectin-4 with puromycin selection
Shipping Temperature
-80°C
Notes
To generate a Nectin-4 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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