The Claudin-9 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 Claudin-9 (NM_020982) driven by an EF1A promoter and a puromycin selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the Claudin-9 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 #
O95484
Background
Claudins are integral membrane proteins and major components of tight junction strands which serve as a physical barrier to prevent solutes and water from passing freely through the paracellular space between epithelial or endothelial cell sheets. Claudin-9 is one of the 27 members of the Claudin family.
Claudin-9 is overexpressed in hepatocellular carcinoma, where it enhances the metastatic potential of hepatocytes via Tyk2/Stat3 signaling. Claudin-9 is also a marker for endometrial cancer, and acts as a receptor for Clostridium perfringens enterotoxin (CPE) during infection by gram-positive Clostridial bacteria. C. perfringens infection causes for food poisoning and gastrointestinal illnesses, via Claudin-9 to toxin-induced gut barrier breakdown.
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 Claudin-9 with puromycin selection
Shipping Temperature
-80°C
Notes
To generate a Claudin-9 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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