pCDH-CuO-MCS-EF1a-CymR-T2A-Puro SparQ All-in-one Cloning and Expression Lentivector

Product Description

With SBI’s SparQ™ Cumate Switch System, you can get inducible gene expression in mammalian cells with a range of cloning and expression lentivectors. This pCDH-CuO-MCS-EF1α-CymR-T2A-Puro SparQ All-in-one Cloning and Expression Lentivector simplifies your SparQ projects by delivering your gene- or miRNA-of-interest with the same vector that delivers CymR. The vector drives your gene-of-interest or miRNA-of-interest with the inducible cumate switch promoter, and constitutively co-expresses CymR and a puromycin resistance marker from an EF1α promoter. Co-expression is mediated by the T2A element.

Simplify your SparQ projects by delivering your gene-of-interest and CymR with the same lentivector – includes co-expression of CymR and a puromycin marker. The SparQ System is characterized by:

• 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
• Versatile—choose from all-in-one formats that co-express CymR and your gene-of-interest, or two-vector systems where CymR is expressed from a different plasmid
• Powerful—suitable for in vivo applications

How it works

The regulatory mechanisms of the bacterial operons cmt and cym have been engineered to regulate gene expression in mammalian cells. In the repressor configuration, regulation is mediated by strong binding of the CymR repressor to the Cumate operator site (CuO), which is downstream of the CMV5 promoter. Addition of cumate (depicted as yellow circle in the graphic scheme below), a non-toxic small molecule inducer, relieves the repression and drives gene activation; in this case, fluorescent proteins (RFP and GFP). Subsequent removal of cumate from the growth media reverts the activation process, rendering gene switches to off mode.

Figure 1. Cumate Switch System

All-in-one Expression Lentivectors of the SparQ™ System

pCDH-CuO-MCS-IRES-GFP-EF1-CymR-T2A-Puro All-in-one inducible IRES lentivector
pCDH-Cuo-MCS-EF1-CymR-T2A-Puro All-in-one cloning and expression lentivector

Two-component SparQ Cloning and Expression Lentivectors

pCDH-CuO-MCS empty cloning vector 
pCDH-CuO-MCS-EF1α-GFP dual promoter lentivector
pCDH-CuO-MCS-EF1α-RFP dual promoter lentivector
pCDH-CuO-MCS-EF1α-GFP+Puro dual promoter and dual marker lentivector
pCDH-CuO-MCS-EF1α-RFP+Puro dual promoter and dual marker lentivector
pCDH-CuO-MCS-T2A-GFP co-inducible T2A lentivector
pCDH-CuO-MCS-T2A-RFP co-inducible T2A lentivector
pCDH-CuO-MCS-IRES-GFP co-inducible IRES lentivector
pCDH-CuO-MCS-IRES-RFP co-inducible IRES lentivector

CymR expression lentivector

CymR expression lentivector pCDH-EF1α-CymR-T2A-Puro as pre-packaged virus

Supporting Data

Tight expression control with low background with the SparQ Cumate Switch System

Figure 2. Get lower background and higher induction with the SparQ Cumate Switch System than other inducible systems.

Figure 3. Gene expression with the SparQ Cumate SwitchSystem can be turned on and off, then on again.

Figure 4. Gene expression with the SparQ Cumate Switch System is titratable, with increasing amounts of cumate inducing a linear increase in gene expression.

Figure 5. With the SparQ System, gene expression can also be titrated by increasing the amount of transduced SparQ lentivirus, even up to 30 MOI.