Overview

Study the balance of T cell dynamics in autoimmunity, inflammation, and cancer

Simplify your studies of T cell dynamics with SBI’s ready-to-use, pre-built Treg and Th17 vectors and cell lines. With the CuO-Mouse Foxp3-IRES-GFP-EF1α-CymR-T2A-Puro Cumate-Inducible Foxp3 Lentivector, you can easily study the effects of different Foxp3 expression levels with SBI’s titratable, cumate-responsive SparQ™ Cumate Switch System. This vector co-expresses the Treg Foxp3 and a GFP marker from the inducible cumate switch promoter, enabling measurement of induction by GFP fluorescence. Co-expression is mediated by an IRES element. The vector also constitutively co-expresses CymR and a puromycin resistance marker from an EF1α promoter, with co-expression mediated by the T2A element.

What’s makes SBI’s SparQ Cumate Switch System so powerful?

With SBI’s SparQ™ Cumate Switch System, you can get inducible gene expression in mammalian cells through the binding of cumate, a non-toxic small molecule, to CymR. Expression levels of your gene-of-interest are tightly controlled and increase with increasing cumate concentration until maximum induction is reached—see as much as a 32-fold increase in gene expression. Even better, induction is reversible, so you can turn expression levels on and off. Delivering negligible background expression in the absence of cumate, the SparQ System delivers controlled levels of gene expression.

• Robust—increase expression up to 32-fold
• Adjustable—tune expression levels by titrating the amount of cumate
• Reversible—turn expression on, then off, then on again
• Powerful—suitable for in vivo applications

How It Works

Meet SBI’s SparQ Cumate Switch System

SBI’s SparQ Cumate Switch System delivers robust, titratable gene expression with low background through three components:

• Cumate, a non-toxic, small-molecule inducer
• CymR, a repressor that binds to cumate operator sequences in the absence of cumate
• SparQ Lentivector that contains an MCS to clone-in your gene-of-interest, the cumate inducible promoter with cumate operator sequences (CuO) upstream of the MCS, and one or more markers

CymR has a high binding affinity for cumate and, as more cumate is added, fewer CymR molecules bind to the CuO sequences in the promoter resulting in increased expression. Exhibiting much lower background expression than similar systems, SBI’s cumate-inducible vectors can provide up to 32-fold induction of gene expression.

Supporting Data

Robust, inducible expression of Foxp3

Figure 1. Robust, inducible expression of Foxp3 as shown by fluorescence of the co-expressed, inducible GFP marker and Western blot analysis. METHODS: The CuO-Mouse Foxp3-IRES-GFP-EF1-CymR-T2A-Puro Lentivector was packaged into lentivirus and transduced into 293FT cells. A stable cell line was established using puromycin selection, and overexpression of Foxp3 induced with cumate. Cells were imaged after seven days. The GFP marker serves as a positive response marker to cumate induction. Cellular proteins were harvested after seven days and western blot analysis performed to test for mFoxp3 protein induction using an anti-Foxp3 antibody, with tubulin as a loading control (tubulin lanes probed with anti-tubulin antibody).

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