Department of Molecular Biology, Umeå University, Umeå, Sweden.
Umeå Centre for Molecular Medicine, Umeå University, Umeå, Sweden.
Nat Commun. 2023 Dec 18;14(1):8410. doi: 10.1038/s41467-023-44177-z.
G protein-coupled receptors (GPCRs) mediate responses to various extracellular and intracellular cues. However, the large number of GPCR genes and their substantial functional redundancy make it challenging to systematically dissect GPCR functions in vivo. Here, we employ a CRISPR/Cas9-based approach, disrupting 1654 GPCR-encoding genes in 284 strains and mutating 152 neuropeptide-encoding genes in 38 strains in C. elegans. These two mutant libraries enable effective deorphanization of chemoreceptors, and characterization of receptors for neuropeptides in various cellular processes. Mutating a set of closely related GPCRs in a single strain permits the assignment of functions to GPCRs with functional redundancy. Our analyses identify a neuropeptide that interacts with three receptors in hypoxia-evoked locomotory responses, unveil a collection of regulators in pathogen-induced immune responses, and define receptors for the volatile food-related odorants. These results establish our GPCR and neuropeptide mutant libraries as valuable resources for the C. elegans community to expedite studies of GPCR signaling in multiple contexts.
G 蛋白偶联受体（GPCRs）介导对各种细胞外和细胞内信号的反应。然而，大量的 GPCR 基因及其显著的功能冗余性使得系统地在体内解析 GPCR 功能具有挑战性。在这里，我们采用基于 CRISPR/Cas9 的方法，在秀丽隐杆线虫的 284 个菌株中破坏了 1654 个 GPCR 编码基因，并在 38 个菌株中突变了 152 个神经肽编码基因。这两个突变文库使化学感受器的去孤儿化和各种细胞过程中神经肽受体的表征成为可能。在单个菌株中突变一组密切相关的 GPCR 允许将功能分配给具有功能冗余的 GPCR。我们的分析确定了一种在缺氧诱导的运动反应中与三个受体相互作用的神经肽，揭示了病原体诱导的免疫反应中的一组调节剂，并定义了与挥发性食物相关气味的受体。这些结果确立了我们的 GPCR 和神经肽突变文库作为秀丽隐杆线虫社区的宝贵资源，以加速在多种情况下研究 GPCR 信号转导。
