CuO-GFP-T2A-Luciferase Cumate-Inducible Dual Reporter Enhanced Episomal Vector (EEV)

An easy-to-produce non-integrating option for inducible GFP and luciferase expression

Product Description

With virtually no limits on insert size (unlike AAV vectors) Enhanced Episomal Vectors (EEVs) are an excellent choice for non-integrating, non-viral gene expression. Because they replicate in synchrony with the host cell, they are stably inherited and can be used for long-lasting expression—up to several months both in vitro and in vivo—without modifying the host genome. Use SBI’s CuO-GFP-T2A-Luciferase EEV Vector for inducible and coordinated expression of GFP and luciferase reporters. The CuO promoter is a tightly-controlled inducible promoter which provides titratable gene expression in response to cumate.

EEV Cumate-Inducible Reporter Constructs In Vitro

The cumate inducible EEV reporter construct features a cumate-inducible promoter driving the expression of GFP-T2A- Luciferase as well as a constitutive EF1 promoter expressing the CymR repressor and a Puromycin selection cassette. The EEV605A-1-SBI cumate switch inducible reporter is silent until the addition of the inducer cumate solution (Cat# QM150A-1-SBI, 10,000x concentration). Cumate was added at a concentration from the 10,000x stock daily at a final concentration in the medium of 300 ug/ml and cells imaged for GFP induction. The GFP induction appeared after 2 days and then expression was detected for 72 days (2.5 months) after the original transfection (fig.1). 

Figure 1. EEV605A-1-SBI construct map and sample data for the minus cumate and the 11 and 72 day expression data.

EEV Cumate-Inducible Reporter Constructs In Vivo

The Cumate inducible EEV605A-1-SBI GFP-T2A-Luciferase reporter plasmid was introduced into test mice (n=3) by using 5 ug plasmid and the HDD injection procedure on Day 0. The water-soluble version of the inducer cumate compound (cat# QM150A-1-SBI) was injected by IP (1.5 mg) per animal to induce EEV expression. The mice were then imaged for luminescence activity through full body scans and liver expression levels quantitated from Day 2 (fig.2) through day 10.

Figure 2. Cumate induction of luciferase mouse images for Day 2 after cumate addition.

For constitutive EEV expression we offer the CAGs-GFP-T2A-Luciferase Dual Reporter Enhanced Episomal Vector (EEV).