Synthetic receptors that mediate antigen-dependent cell responses are transforming therapeutics, drug discovery, and basic research. However, established technologies such as chimeric antigen receptors (CARs) can only detect immobilized antigens, have limited output scope, and lack built-in drug control. Here, we engineer synthetic G protein-coupled receptors (GPCRs) capable of driving a wide range of native or nonnative cellular processes in response to user-defined antigen. We achieve modular antigen gating by engineering and fusing a conditional auto-inhibitory domain onto GPCR scaffolds. Antigen binding to a fused nanobody relieves auto-inhibition and enables receptor activation by drug, thus generating Programmable Antigen-gated G protein-coupled Engineered Receptors (PAGERs). We create PAGERs responsive to more than a dozen biologically and therapeutically important soluble and cell surface antigens, in a single step, from corresponding nanobody binders. Different PAGER scaffolds permit antigen binding to drive transgene expression, real-time fluorescence, or endogenous G protein activation, enabling control of cytosolic Ca2+, lipid signaling, cAMP, and neuronal activity. Due to its modular design and generalizability, we expect PAGER to have broad utility in discovery and translational science.

中文翻译：

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用于细胞行为可编程传感和控制的合成 G 蛋白偶联受体

介导抗原依赖性细胞反应的合成受体正在改变治疗方法、药物发现和基础研究。然而，嵌合抗原受体（CAR）等现有技术只能检测固定化抗原，输出范围有限，并且缺乏内置药物控制。在这里，我们设计了合成的 G 蛋白偶联受体 (GPCR)，能够驱动各种天然或非天然细胞过程来响应用户定义的抗原。我们通过设计条件自动抑制结构域并将其融合到 GPCR 支架上来实现模块化抗原门控。抗原与融合纳米抗体的结合可解除自身抑制，并通过药物激活受体，从而产生可编程抗原门控 G 蛋白偶联工程受体 (PAGER)。我们通过相应的纳米抗体结合剂一步创建了对十多种具有生物学和治疗学意义的可溶性和细胞表面抗原有反应的寻呼机。不同的 PAGER 支架允许抗原结合来驱动转基因表达、实时荧光或内源性 G 蛋白激活，从而能够控制胞质 Ca2+、脂质信号传导、cAMP 和神经元活性。由于其模块化设计和通用性，我们预计 PAGER 在发现和转化科学中具有广泛的用途。