ABSTRACT
This chapter focuses on the functions of Orai/Stim proteins studied in immunity in transgenic animal models and describes the strategies for making tissue-specific Orai1, Stim1, and Stim2 gene knockout mouse. Three main strategies for the gene targeting have been utilized so far, for example, conventional gene targeting, conditional gene targeting, and gene trapping. A gene-trap vector in its most basic version contains a splice-acceptor sequence at the 5' end, a reporter gene, a selectable marker gene, and a polyadenylation signal sequence at the 3' end. Orai1 protein, encoded by the Orai1 gene located on chromosome 5 in the mouse, is a small and widely expressed tetraspanning membrane glycoprotein, whose N- and C-termini are both located in the cytoplasm. Stim1 protein, encoding the Stim1 gene located on chromosome 7 in the mouse, is a type I transmembrane protein predominantly localized in the endoplasmic reticulum membrane and expressed broadly.
