AAV with CAG promoter driven roGFP1

This AAV expresses roGFP1 driven by an ubiquitous CAG promoter.

The roughly 1.8Kb CAG promoter (also known as CBA promoter or CAGGS promoter) is a strong synthetic promoter frequently used to drive high levels of gene expression in mammalian cells. The CAG promoter is composed of the following regulatory elements: (C) cytomegalovirus (CMV) early enhancer element; (A) the promoter region, the first exon, and the first intron of chicken beta-Actin gene, and (G) the splice acceptor of the rabbit beta-Globin gene. Like the EF1a promoter, the CAG promoter is commonly used as an alternative for the CMV promoter, from which the expression is decreased due to methylation/silencing. In many cell types tested, the CAG and EF1a promoters give much higher levels of expression than other commonly used cellular promoters such as the UBC and PGK promoters.

roGFP1 is a reduction-oxidation sensitive green fluorescent protein used as a biosensor. Two cysteines were introduced into the beta-barrel structure of GFP; the oxidation state, reduced dithiol or the oxidized disulfide, of the engineered thiols determines the fluorescent properties of the sensor. roGFPs (roGFP1 and roGFP2), have two fluorescent excitation maxima at about 400 and 490 nm, respectively and display rapid and reversible ratiometric changes in fluorescence in response to changes in redox potential in vitro and in vivo.