Gt(ROSA)26Sor^{tm1(CAG-Eos*/Calm*)Mrc}

The targeting construct contains a CAG promoter followed by a loxP-Stop-loxP-CaMPARI2::attB-PGK-FRT-Neo-pA-attP integrated into intron 1 of the Gt(ROSA)26Sor locus. CaMPARI (Calcium Modulated Photoactivatable Ratiometric Integrator) is a photoconvertible protein construct derived from the stony coral protein Eos. CaMPARI and irreversibly photoconverts from green to red in the presence of violet light and high intracellular calcium. The CaMPARI protein construct includes a calcium binding calmodulin with a calmodulin/Ca<2+> binding peptide (M13) fused to either end of the protein making the photoconversion calcium dependent. Calcium binding affinity variants of the original CaMPARI (CaMPARI1) fluorescent protein were identified (CaMPARI1_W391F-V398L_Q142V-F198Y-C202T-L217I-N345S) and incorporated in CaMPARI2 via targeted mutagenesis of amino acids around the fluorescent protein chromophore and at the protein interface between the fluorescent protein and the calcium-sensitive domains. (J:101977)