Pooled functional genomic screens for intracellular calcium effectors

Abstract

Cytoplasmic calcium transients relay cellular signals on timescales from milliseconds to hours, and the dynamic nature of calcium signals has slowed functional genomic screening for cellular calcium effectors. Here, we present a new strategy to identify calcium handling genes via a pooled knockdown employing the calcium-sensitive photo-switchable fluorescent protein, CaMPARI. This assay, cal-Seq, enabled identification of regulators for both cellular desensitization and histamine induced calcium signaling including GPR99, a leukotriene binding receptor, with implications in asthma treatment.