Tg(Acta2-RHO/ADRA1,-lacZ)B6-1Mik

The transgenic construct is made using a C57BL/6J mouse bacterial artificial chromosome (BAC) RP23-370F21 containing the entire actin alpha 2 locus (Acta2) as well as ~92 kbp of 5' flanking sequences (including the complete Fas locus) and ~68 kbp of 3' flanking sequences (including the complete Stambpl1 locus). Using homologous recombination/BAC recombineering, a 6170 bp Opto alpha;1AR-IRES-lacZ-pA construct is inserted into the ATG start site of the BAC Acta2 gene (replacing the initiation codon in exon 2). No other loci on the BAC are altered. The 11.6 kbp BAC vector (pBACe3.6) contains a chloramphenicol selection cassette. The Opto alpha 1AR fusion protein is a Gt-coupled bovine rhodopsin (RHO) sequence modified to have its intracellular regions (including carboxy-terminal domains) replaced with those from the Gq-coupled human ADRA1A. Founder line B6-1 has high sensor expression in purified smooth muscle. (J:314125)