The Trop2 (Trophoblast cell-surface antigen 2) Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These viruses transduce cells with Macaca fascicularis (also known as crab-eating macaque or cynomolgus monkey) Trop2 (XP_005543292.2) driven by an EF1A promoter. The lentiviruses also transduce a puromycin selection gene (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the cynomolgus Trop2 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 in HEK293T cells in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.
Background
Trophoblast cell-surface antigen 2 (Trop-2, also known as tumor-associated calcium signal transducer 2, TACSTD2, GA733-1 or M1S1), is a cell surface glycoprotein involved in stem-cell like functions. It is overexpressed in a variety of solid cancers compared to normal cells, and links with patient prognosis. Through a variety of signaling pathways, Trop-2 regulates cancer growth and metastasis. The use of antibodies, antibody-fusion proteins, small molecule inhibitors and other strategies directed at Trop-2 has resulted in promising results. Its use as biomarker and target in cancer therapy shows its importance in the field.
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.
To generate a Trop2 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 a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after 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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