Illumination of TAAR2 Location, Function and Regulators

项目摘要 本提案是应RFA-RM-22-024号文件的要求编写的，该文件邀请了 对G蛋白偶联受体、离子通道和蛋白激酶研究不足“。我们选择研究痕量胺 相关受体2(TAAR2)是一种未被充分研究的GPCR，是一种符合IDG条件的蛋白。 TAAR2在2001年被发现是一种gpr基因，并被推测参与了神经学和 精神疾病；然而，在理解TAAR2和TAAR2的作用方面进展甚微 创造实验工具。为了解决目前的知识差距，我们将研究TAAR2细胞和 利用人SH-SY5Y细胞和啮齿类动物脑的亚细胞定位、运输和信号转导。 此外，该方案还将CRISPR/Cas9介导的同源定向修复机制应用于标签 TAR2基因在SH-SY5Y细胞中的表达，从而通过蛋白质组学方法揭示与TAAR2相互作用的蛋白质网络。 从该计划中学到的知识和产生的产品将为IDG计划增加价值，并 重要的是，为未来在人类疾病动物模型中研究TAAR2奠定了基础。

Project Summary This proposal is prepared in response to RFA-RM-22-024, which invites “Pilot projects investigating understudied G protein-coupled receptors, ion channels and protein kinases”. We choose to study trace amine associated receptor 2 (TAAR2) which is an understudied GPCR and an IDG-eligible protein under this FOA. TAAR2 was discovered as a GPCR in 2001 and has been speculated to be involved in neurological and psychiatric diseases; however, little progress has been made towards understanding the action of TAAR2 and creating experimental tools. To address the current knowledge gap, we will investigate TAAR2 cellular and subcellular localization, trafficking and signal transduction using human SH-SY5Y cells and rodent brains. Further, this proposal will also apply CRISPR/Cas9-mediated homology-directed repair mechanism to tag Taar2 gene in SH-SY5Y cells so that TAAR2-interacting protein network can be revealed through proteomics. Knowledge learned and products generated from this proposal will add value to the IDG program and importantly, lay ground work for future investigation of TAAR2 in animal models of human diseases.