AAV with hRPE(0.8) promoter driven roGFP1

This AAV expresses roGFP1 driven by a retina hRPE(0.8) promoter.

The RPE65 gene encodes a protein of the retinal pigment epithelium (RPE) with a key role in the cycling of retinoids. It was shown that a RPE promoter of about 0.8 Kb was sufficient to direct high RPE-specific expression in transgenic mice. The 1.5Kb version of the promoter hRPE(1.5), also gives RPE-specific expression, but the expression level is not as high as the 0.8Kb promoter. This is probably due to a negative regulatory element(s) present in the 1.5Kb promoter.

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.