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As a first and second messenger, calcium (Ca2+) regulates numerous biological processes. Calcium homeostasis is modulated by intracellular Ca2+ stores, membrane channels, pumps, and/or receptors. Numerous diseases have been reported to be related to mishandling of calcium. Calcium signaling and calcium dynamics are mediated by major calcium stores and extracellular receptors, especially by calcium-sensing receptor (CaSR). In this chapter, we first introduce common properties for calcium-binding sites in proteins. We will then review efforts in addressing a pressing need to integrate Ca2+ signaling intracellularly and extracellularly, using a novel class of genetically encoded Ca2+ indicators (GECIs) with rapid kinetics. We describe a general strategy to rationally design a new class of GECIs, with a single Ca2+-binding site and fast kinetics, by tuning rapid protein dynamics to modulate the chromophore …