Tg(tetO-CHRM3*)1Blr

The Tet-responsive P promoter which consists of seven direct repeats of the 36-bp sequence each containing the 19-bp tet operator sequence (tetO) is just upstream of a minimal human cytomegalovirus (CMV) promoter. This is upstream of a hemagglutinin epitope tag and a Gq-coupled human M3 muscarinic receptor. The wild-type human muscarinic 3 receptor sequence was modified via site-directed mutagenesis to harbor two amino acid substitutions (Y149C and A239G) that abolish receptor affinity for the native ligand, acetylcholine (ACh), but allow receptor binding and subsequent activation by the small drug-like molecule clozapine-N-oxide (CNO). (J:154952)