Functional Genomics of G Protein-Coupled Receptors

G蛋白偶联受体(GPCRs)是哺乳动物细胞感知和反应能力的中枢介质 到他们的环境。813个人类GPCRs是最大的一类膜受体，是中枢 细胞生理调节剂，是美国食品和药物管理局约34%的目标 (FDA)批准的药品和约60%的处方。在这里，我们将利用DNA合成的最新进展， 基因组编辑、下一代DNA测序和信息学开发了一个平台，建立了数千个 GPCRs的单个突变并在一种新的可以完成的测试中实验表征它们的影响 在人类细胞系中以一种简单的混合形式。我们将开发方法来描述这些 突变文库在功能上通过GPCRs信号传递的两条主要途径发出信号，即环状 AMP和钙信号通路。这些配置文件将使我们能够更好地理解功能 GPCR基因变异的后果以及它们可能如何影响疾病和药物反应。 此外，对这些突变图谱的分析将使我们深入了解如何针对这些受体 制造新药。这里开发的技术应该广泛和直接适用于广大 大多数GPCRs，以及其他重要的药物靶点，其作用可以通过 人类细胞系中的转录记者。

G protein-coupled receptors (GPCRs) are central mediators of mammalian cells’ ability to sense and respond to their environment. The 813 human GPCRs are the largest class of membrane receptors, are central mediators of cell physiology, and are the target of ~34% of all U.S. Food and Drug Administration (FDA)-approved drugs and ~60% of prescriptions. Here we will leverage recent advances in DNA synthesis, genome editing, next-generation DNA sequencing, and informatics develop a platform to build thousands of individual mutations to GPCRs and experimentally characterize their effects in a novel assay that can be done in a simple pooled format in human cell lines. We will develop methodologies to characterize how these mutational libraries functionally signal through the two major pathways that GPCRs signal through, the cyclic AMP and calcium signaling pathways. The profiles will give us the ability to better understand the functional consequences of genetic variation in GPCRs and how they might be impacting diseases and drug responses. In addition, analysis of these mutational profiles will give us insights on how to target these receptors for making new drugs. The technologies developed here should be broadly and directly applicable to the vast majority of GPCRs, as well as other important classes of drug targets where function can be assessed by transcriptional reporters in human cell lines.