AAV with tMCK promoter driven Mito-roGFP1

This AAV expresses Mito-roGFP1 driven by a muscle tMCK promoter.

tMCK is constructed by ligating a triple tandem copies of mouse MCK enhancer ( about200 bps each) to the ~100bp mouse MCK basal promoter.). This promoter shows extremely sensitive tissue-specificity: (1) In differentiated C2C12 myotubes, the tMCK promoter is 30-50 fold stronger than the Enh358MCK promoter and 10-20 fold stronger than the CMV promoter; (2) In muscle tissue, tMCK promoter is 15-20 fold stronger than the Enh358MCK promoter, and 3-5 fold stronger than the CMV promoter; (3) Strong tissue-specificity: in liver tissue, the expression level from tMCK promoter is only about 0.3-0.5% of that of the CMV promoter.

The mitochondrial targeting sequence of human pyruvate dehydrogenase alpha 1 (PDHA1) is fused in-frame with the redox-sensitive green fluorescent protein (roGFP1). The Mito-roGFP1 is a biosensor for investigating the mitochondrial reduction-oxidation potential.