Tg(Acta2-COP4*L132C,-lacZ)B8-3Mik

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 5640 bp COP4-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 light-activated, non-specific ion channel chlamyopsin 4 (COP4 or ChR2) cDNA sequence used here encodes the first 309 amino acids of green alga Chlamydomonas reinhardtii channelrhodopsin-2 that has been modified with an L132C amino acid substitution (CTC-to-TGC) designed to cause enhanced Ca2+ permeability. Founder line B8-3 has high sensor expression in smooth muscle. (J:314125)