CAGs-GFP-T2A-Luciferase Dual Reporter Enhanced Episomal Vector (EEV)

An easy-to-produce non-integrating option for constitutive 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 CAGs-GFP-T2A-Luciferase EEV Vector  for constitutive, CAG-driven coordinated expression of GFP and luciferase reporters in most cell types, including primary cells and stem cells.

EEV Constitutive Reporter Constructs In Vitro

The constitutive EEV reporter construct was transfected into HEK293T cells and the GFP transgene expression monitored over a period of several weeks. HEK293T cells in a 24-well culture dish were transfected once with 0.5 ug of EEV604A-1-SBI reporter plasmid DNA. The GFP signals were then monitored over a period of several weeks (fig.1). 

Figure 1. EEV604A-1-SBI construct map and sample cell images from 2 days and 11 days after transfections

EEV Constitutive Reporter Constructs In Vivo

The constitutive EEV604A-1-SBI CAGs-GFP-T2A-Luciferase construct (8 ug plasmid DNA) was introduced into test mice through hydrodynamic tail vein injection (HDD). This procedure leads to high plasmid DNA transfection of the livers of mice in vivo. The test mice (n=3) were imaged for body luminescence in the liver area post HDD from Day 1 up until Day 80. The results show that robust EEV-expressed luciferase expression is readily detectable at very high levels through day 40 (fig.2). 

Figure 2. Luciferase quantitative data for the test mice in graphical form as well as bioluminescent animal imaging data.

For inducible EEV expression we offer the CuO-GFP-T2A-Luciferase Cumate-Inducible Dual Reporter Enhanced Episomal Vector (EEV).