Sphkap^tm1.1Nina

An mCherry-poly(A)-loxp-Neo-loxp construct was inserted into exon 1 of the gene. The loxP-flanked neomycin selection cassette was removed via Cre-mediated recombination. mCherry is expressed under the control of the endogenous promoter. RT-PCR on samples from islets showed that no endogenous transcript is expressed from this allele. Western blot analysis confirmed the absence of protein expression in isolated pancreatic islets from homozygous mutant mice. Immunohistochemical analysis showed that mCherry is detected only in beta-cells, and not in alpha-cells from mutant mice. Incubator two-photon excitation microscopy confirmed that mCherry signal is strongly detected in single beta-cells of mutant mice, but not in wild-type mice. (J:272327)