The CEACAM5 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 CEACAM5 (NM_004363.6) driven by an EF1A promoter and a puromycin selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the CEACAM5 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 #
P06731
Background
Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5, also known as CD66e) is a cell surface glycoprotein that serves as a cell adhesion protein. It has been used as a clinical biomarker to detect liver metastasis from gastrointestinal cancers and to predict gastrointestinal cancer relapse. CEACAM5 was recently identified as a potential target antigen for CAR T-cell therapy. Additionally, CEACAM5 may be involved in the inhibition of cell differentiation, apoptosis, and cell polarity.
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 CEACAM5 with puromycin selection
Shipping Temperature
-80°C
Notes
To generate a CEACAM5 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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