G-蛋白偶联受体(GPCRs)构成跨膜受体的最大家族,并且是大约三分之一的FDA批准的药物的靶标,因为它们参与几乎所有的生理过程。GPCR信号通过异源三聚体G蛋白复合物和β-抑制素的激活而发生,两者都作为换能器,导致不同的细胞反应。尽管乍一看似乎很简单,越来越多的证据表明,任何一个换能器的激活都不是一个简单的过程,作为单个分子的刺激具有激活多个信号分支的潜力。GPCR信号的复杂性来自G蛋白偶联选择性方面,有偏见的信号,通路间串扰,和可变的分子修饰产生这些不同的信号模式。在CRISPR-Cas9基因组编辑的最新发展之前,与这些信号传导方面有关的许多问题仍未得到解答。这种基因组编辑技术提供了长期消除G蛋白亚基表达的机会,β-抑制素,GRKs,以及GPCR途径中的许多其他信号节点。这里,我们回顾了在揭示GPCR信号传导机制的分子细节的实验环境中使用CRISPR-Cas9衍生的敲除(KO)细胞的实用性.举几个例子,KO细胞揭示了β-抑制素在ERK激活中的作用,Gα蛋白选择性,基于GRK的GPCR监管,还有更多,因此验证了其在GPCR研究中的广泛适用性。意义陈述这篇综述强调了GPCR效应KO细胞在解剖GPCR信号网络的复杂调节机制中的实际应用。目前可用的细胞系,随着KO细胞系在不同细胞类型中的积累,为系统地阐明GPCR信号传导调控提供了宝贵的资源。鉴于GPCR信号与许多疾病的关联,揭示基于系统的信号图谱对于推进针对特定疾病的新型药物的开发至关重要.
G-protein-coupled receptors (GPCRs) compose the largest family of transmembrane receptors and are targets of approximately one-third of Food and Drug Administration-approved drugs owing to their involvement in almost all physiologic processes. GPCR signaling occurs through the activation of heterotrimeric G-protein complexes and β-
arrestins, both of which serve as transducers, resulting in distinct cellular responses. Despite seeming simple at first glance, accumulating evidence indicates that activation of either transducer is not a straightforward process as a stimulation of a single molecule has the potential to activate multiple signaling branches. The complexity of GPCR signaling arises from the aspects of G-protein-coupling selectivity, biased signaling, interpathway crosstalk, and variable molecular modifications generating these diverse signaling patterns. Numerous questions relative to these aspects of signaling remained unanswered until the recent development of CRISPR genome-editing technology. Such genome editing technology presents opportunities to chronically eliminate the expression of G-protein subunits, β-
arrestins, G-protein-coupled receptor kinases (GRKs), and many other signaling nodes in the GPCR pathways at one\'s convenience. Here, we review the practicality of using CRISPR-derived knockout (KO) cells in the experimental contexts of unraveling the molecular details of GPCR signaling mechanisms. To mention a few, KO cells have revealed the contribution of β-
arrestins in ERK activation, Gα protein selectivity, GRK-based regulation of GPCRs, and many more, hence validating its broad applicability in GPCR studies. SIGNIFICANCE STATEMENT: This review emphasizes the practical application of G-protein-coupled receptor (GPCR) transducer knockout (KO) cells in dissecting the intricate regulatory mechanisms of the GPCR signaling network. Currently available cell lines, along with accumulating KO cell lines in diverse cell types, offer valuable resources for systematically elucidating GPCR signaling regulation. Given the association of GPCR signaling with numerous diseases, uncovering the system-based signaling map is crucial for advancing the development of novel drugs targeting specific diseases.