DLL3 (Delta-like protein 3) Lentivirus 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 particles contain human DLL3 (NM_016941.3) driven by a CMV promoter and a puromycin selection marker (Figure 1).
Figure 1. Schematic of the lenti-vector used to generate the DLL3 Lentivirus.
●
Product Data Gallery
Expression of human DLL3 in CHO cells transduced with DLL3 lentiviruses.
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
DLL3, also known as delta like ligand three, is a Notch ligand characterized by a DSL domain, transmembrane region, and a series of EGF repeats. Notch ligands can participate in trans-interactions (interaction with Notch receptor on a different cell) and cis interactions (interaction with Notch receptor within the same cell) to activate or inhibit Notch signaling, respectively. DLL3 exclusively functions to inhibit Notch signaling through cis inhibition. While DLL3 expression is limited in healthy tissue, high expression levels of DLL3 are found in various cancers including small cell lung cancer (SCLC), where it plays an oncogenic role. Relieving DLL3-mediated inhibition of Notch signaling may serve as a therapeutic avenue, with drugs being developed to target DLL3 as a possible lung cancer therapy (example: rovalpituzumab tesirine).
References
Chapman G, et al., 2011 Hum Mol Genet. 20(5):905-16
Ladi E, et al., 2005 J Cell Biol. 170 (6): 983–992
Kunnimalaiyaan M, et al., 2007 The oncologist. 12(5):535-42
Owen D, et al., 2019 J Hematol Oncol. 12(1): 61
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
Expression of human DLL3 in cells of interest.
Generate cell pools or stable cell lines expressing human DLL3 following puromycin selection
Shipping Temperature
-80°C
Notes
To generate a DLL3 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, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, followed by clonal selection.
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.