Tg(Acta2-GCaMP-ER2)B25-6Mik

Acta2-GCaMP-ER2 transgenic mice (also called Acta2-GCaMP-ER or Acta2-GCaMP-ER2) were designed in the laboratory of Dr. Michael I. Kotlikoff (Cornell University) as part of Cornell/National Heart Lung Blood Resource for Optogenetic Mouse Signaling (CHROMus). The ~175 kbp C57BL/6J mouse bacterial artificial chromosome (BAC) RP23-370F21 was obtained; 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 2344 bp GCaMP-ER2-SV40pA construct (details below) was inserted into the ATG start site of the BAC Acta2 gene (replacing the initiation codon of Acta2 in exon 2). No other loci on the BAC were altered. The 11,612 bp BAC vector (pBACe3.6) contained a chloramphenicol selection cassette. (J:101977)