Gt(ROSA)26Sor^{tm87(CAG-gltI/GFP*)Hze}

The Rosa-CAG targeting vector CAG-LSL-iGluSnFR-WPRE-bGHpA was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. The targeting vector was designed with a floxed-STOP cassette, the iGluSnFr coding sequence (details below), a WPRE sequence (to enhance mRNA stability), a BGH polyA. The iGluSnFr fusion gene is constructed from the Escherichia coli gltI gene and circularly permutated green fluorescent protein (cpGFP). Specifically, iGluSnFr has a 21 aa IgG K-leader sequence, the gltI aa 1-253, the LILV linker, the cpGFP (aa 148-239, an 18 bp flexible linker, and aa 1-147), the L2NP linker, the gltI aa 254-279, an 11 aa c-Myc epitope and a 49 aa PDGR sequence (PDGR aa 513-561). iGluSnFr is an extremely fast and sensitive glutamate indicator with a large signal-to-noise ratio, and kinetics appropriate for in vivo imaging. It is a Cre-dependent reporter with a glutamate indicator function. (J:219930)