Natural display of nuclear-encoded RNA on the cell surface and its impact on cell interaction
Natural display of nuclear-encoded RNA on the cell surface and its impact on cell interaction
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DOI：
10.1186/s13059-020-02145-6
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发表时间：
2020-09-10
期刊：
影响因子：
12.3
通讯作者：
Zhong, Sheng
中科院分区：
文献类型：
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作者：
Huang, Norman;Fan, Xiaochen;Zhong, Sheng
与蛋白质、聚糖和脂质相比，对细胞表面RNA的了解要少得多。我们开发了一系列技术来测试任何稳定附着于细胞表面并暴露于细胞外空间的核编码RNA，以下称为膜相关细胞外RNA（maxRNA）。结果我们开发了一种称为Surface-seq的技术来选择性地测序maxRNA，并通过RNA荧光原位杂交验证了两个Surface-seq鉴定的maxRNA。为了测试maxRNA的细胞类型特异性，我们使用反义寡核苷酸与暴露在人外周血单核细胞（PBMC）表面上的单链转录物杂交。将该策略与成像流式细胞术、单细胞RNA测序和maxRNA测序相结合，我们将单核细胞鉴定为maxRNA+ PBMC的主要类型，并优先考虑11种候选maxRNA用于功能测试。细胞外应用FNDC 3和CTSS转录本的反义寡核苷酸抑制单核细胞与血管内皮细胞的粘附总的来说，这些数据突出了maxRNA作为细胞表面的功能组分，表明RNA在细胞-细胞和细胞-环境相互作用中的作用扩大。
Background Compared to proteins, glycans, and lipids, much less is known about RNAs on the cell surface. We develop a series of technologies to test for any nuclear-encoded RNAs that are stably attached to the cell surface and exposed to the extracellular space, hereafter called membrane-associated extracellular RNAs (maxRNAs). Results We develop a technique called Surface-seq to selectively sequence maxRNAs and validate two Surface-seq identified maxRNAs by RNA fluorescence in situ hybridization. To test for cell-type specificity of maxRNA, we use antisense oligos to hybridize to single-stranded transcripts exposed on the surface of human peripheral blood mononuclear cells (PBMCs). Combining this strategy with imaging flow cytometry, single-cell RNA sequencing, and maxRNA sequencing, we identify monocytes as the major type of maxRNA+ PBMCs and prioritize 11 candidate maxRNAs for functional tests. Extracellular application of antisense oligos ofFNDC3BandCTSStranscripts inhibits monocyte adhesion to vascular endothelial cells. Conclusions Collectively, these data highlight maxRNAs as functional components of the cell surface, suggesting an expanded role for RNA in cell-cell and cell-environment interactions.