Mahajan Divyanshu, Chia Hui Min, Lu Lei
School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551, Singapore.
Sci Rep. 2025 Jul 24;15(1):26980. doi: 10.1038/s41598-025-11124-5.
Small G protein ARL13B localizes to the cilium and plays essential roles in cilium biogenesis, organization, trafficking, and signaling. Here, we established multiple ARL13B knockout cell lines using the CRISPR/Cas9 system. Surprisingly, all our cell lines lost their cilia completely, in contrast to the reported short cilium and reduced ciliogenesis phenotype. We found that multiple regions of ARL13B are necessary for a complete rescue. Additionally, we found that ARL13B knockout cells also lost their response to SMO-mediated hedgehog stimulation. Our work demonstrates the critical requirement of ARL13B for ciliogenesis and hedgehog signaling, at least in cultured cells, and suggests that ARL13B plays a more crucial role in ciliary function than previously understood.
小G蛋白ARL13B定位于纤毛，在纤毛的生物发生、组织、运输和信号传导中发挥重要作用。在此，我们使用CRISPR/Cas9系统建立了多个ARL13B基因敲除细胞系。令人惊讶的是，与报道的短纤毛和纤毛发生减少的表型相反，我们所有的细胞系都完全失去了纤毛。我们发现ARL13B的多个区域对于完全挽救是必需的。此外，我们发现ARL13B基因敲除细胞也失去了对SMO介导的刺猬信号刺激的反应。我们的工作证明了ARL13B对于纤毛发生和刺猬信号传导的关键需求，至少在培养细胞中是如此，并表明ARL13B在纤毛功能中发挥着比以前所理解的更关键的作用。
