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GAP, an aequorin-based fluorescent indicator for imaging Ca2+ in organelles Rodríguez García, Arancha Rojo Ruiz, Jonatan Navas Navarro, Paloma Aulestia Araya, Francisco Javier Gallego Sandin, Sonia García-Sancho Martín, Francisco Javier Alonso Alonso, María Teresa Calcio Radiología Producción Científica Genetically encoded calcium indicators allow monitoring subcellular Ca2+ signals inside organelles. Most genetically encoded calcium indicators are fusions of endogenous calcium-binding proteins whose functionality in vivo may be perturbed by competition with cellular partners.We describe here a novel family of fluorescent Ca2+ sensors based on the fusion of two Aequorea victoria proteins, GFP and apo-aequorin (GAP). GAP exhibited a unique combination of features: dual-excitation ratiometric imaging, high dynamic range, good signal-to-noise ratio, insensitivity to pH and Mg2+, tunable Ca2+ affinity, uncomplicated calibration, and targetability to five distinct organelles. Moreover, transgenic mice for endoplasmic reticulum-targeted GAP exhibited a robust long-term expression that correlated well with its reproducible performance in various neural tissues. This biosensor fills a gap in the actual repertoire of Ca2+ indicators for organelles and becomes a valuable tool for in vivo Ca2+ imaging applications. 2014-09-30T18:04:21Z 2014-09-30T18:04:21Z 2014 info:eu-repo/semantics/article Proceedings of the National Academy of Sciences, Febrero 2014, vol. 111, n. 7, p. 2584-2589 1091-6490 http://uvadoc.uva.es/handle/10324/6350 10.1073/pnas.1316539111 2584 7 2589 Proceedings of the National Academy of Sciences 111 eng Attribution-NonCommercial-NoDerivatives 4.0 International info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf National Academy of Sciences https://uvadoc.uva.es/bitstream/10324/6350/6/GARCIA-SANCHO%207.pdf.jpg Hispana TEXT http://creativecommons.org/licenses/by-nc-nd/4.0/ UVaDOC. Repositorio Documental de la Universidad de Valladolid http://uvadoc.uva.es/handle/10324/6350