The STIM-Orai Pathway: Light-Operated Ca²⁺ Entry Through Engineered CRAC Channels

Ca²⁺ signals regulate a plethora of cellular functions that include muscle contraction, heart beating, hormone secretion, lymphocyte activation, gene expression, and metabolism. To study the impact of Ca²⁺ signals on biological processes, pharmacological tools and caged compounds have been commonly applied to induce fluctuations of intracellular Ca²⁺ concentrations. These conventional approaches, nonetheless, lack rapid reversibility and high spatiotemporal resolution. To overcome these disadvantages, we and others have devised a series of photoactivatable genetically encoded Ca²⁺ actuators (GECAs) by installing light sensitivities into a bona fide highly selective Ca²⁺ channel, the Ca²⁺ release-activated Ca²⁺ (CRAC) channel. Store-operated CRAC channel serves as a major route for Ca²⁺ entry in many cell types. These GECAs enable remote and precise manipulation of Ca²⁺ signaling in both excitable and non-excitable cells. When combined with nanotechnology, it becomes feasible to wirelessly photo-modulate Ca²⁺-dependent activities in vivo. In this chapter, we briefly review most recent advances in engineering CRAC channels to achieve optical control over Ca²⁺ signaling, outline their design principles and kinetic features, and present exemplary applications of GECAs engineered from CRAC channels.