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Natural display of nuclear-encoded RNA on the cell surface and its impact on cell interaction.

  • Author(s): Huang, Norman
  • Fan, Xiaochen
  • Zaleta-Rivera, Kathia
  • Nguyen, Tri C
  • Zhou, Jiarong
  • Luo, Yingjun
  • Gao, Jie
  • Fang, Ronnie H
  • Yan, Zhangming
  • Chen, Zhen Bouman
  • Zhang, Liangfang
  • Zhong, Sheng
  • et al.
Abstract

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 of FNDC3B and CTSS transcripts 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.

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