Gt(ROSA)26Sor^{tm1(CAG-GCaMP6f)Khakh}

The targeting vector is designed with (from 5' to 3') with a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), an FRT site, a loxP-flanked STOP cassette, the GCaMP6f coding sequence with N-terminal Lck tag (described below), a WPRE, a BGH polyA signal, and an attB/attP-flanked PGK-frt-Neo-polyA cassette. Lck-GCaMP6f is a GCaMP6 fast variant calcium indicator with an N-terminal Lck-tag that targets GCaMP6f to the plasma membrane. In the absence of calcium binding, dim EGFP fluorescence is expected as there is a pore from the outside of its barrel into the chromophore. Upon calcium binding, this pore becomes occupied and fluorescence is increased. (J:249928)