Spatial imaging of glycoRNA in single cells with ARPLA.
Spatial imaging of glycoRNA in single cells with ARPLA.
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DOI：
10.1038/s41587-023-01801-z
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发表时间：
2023-05
影响因子：
46.9
通讯作者：
Yuan Ma;Weijie Guo;Quanbing Mou;Xiangli Shao;Mingkuan Lyu;Valeria Garcia;Linggen Kong;Whitney Lewis;Carson Ward;Zhenglin Yang;Xingxin Pan;S. S. Yi-S.;Yi Lu
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文献类型：
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作者：
Yuan Ma;Weijie Guo;Quanbing Mou;Xiangli Shao;Mingkuan Lyu;Valeria Garcia;Linggen Kong;Whitney Lewis;Carson Ward;Zhenglin Yang;Xingxin Pan;S. S. Yi-S.;Yi Lu
由于缺乏可视化方法，人们对糖基化RNA的生物学作用知之甚少。糖基化RNA是最近发现的一类糖基化分子。我们报道了唾液酸适配子和RNA原位杂交介导的邻近连接试验(ARPLA)以高灵敏度和选择性显示单细胞中的糖RNAs。只有当糖和RNA的双重识别触发原位连接，然后滚动循环扩增互补DNA，通过结合荧光团标记的寡核苷酸产生荧光信号时，ARPLA的信号输出才会发生。利用ARPLA，我们检测了糖RNA在细胞表面的空间分布及其与脂筏的共存，以及糖RNA通过SNARE蛋白介导的分泌性胞吐在细胞内的运输。对乳腺细胞系的研究表明，表面糖核酸与肿瘤的恶性程度和转移呈负相关。对糖RNA与单核细胞-内皮细胞相互作用关系的研究表明，在免疫应答过程中，糖RNA可能介导细胞-细胞相互作用。
Little is known about the biological roles of glycosylated RNAs (glycoRNAs), a recently discovered class of glycosylated molecules, because of a lack of visualization methods. We report sialic acid aptamer and RNA in situ hybridization-mediated proximity ligation assay (ARPLA) to visualize glycoRNAs in single cells with high sensitivity and selectivity. The signal output of ARPLA occurs only when dual recognition of a glycan and an RNA triggers in situ ligation, followed by rolling circle amplification of a complementary DNA, which generates a fluorescent signal by binding fluorophore-labeled oligonucleotides. Using ARPLA, we detect spatial distributions of glycoRNAs on the cell surface and their colocalization with lipid rafts as well as the intracellular trafficking of glycoRNAs through SNARE protein-mediated secretory exocytosis. Studies in breast cell lines suggest that surface glycoRNA is inversely associated with tumor malignancy and metastasis. Investigation of the relationship between glycoRNAs and monocyte–endothelial cell interactions suggests that glycoRNAs may mediate cell–cell interactions during the immune response.