SUGAR-Seq enables simultaneous detection of glycans, epitopes and the transcriptome in single cells [3'RNA-seq]. SUGAR-Seq enables simultaneous detection of glycans, epitopes and the transcriptome in single cells [3'RNA-seq]

Multi-modal single cell RNA sequencing enables the precise mapping of transcriptional and phenotypic features of cellular differentiation states, but does not allow for simultaneous integration of critical post-translational modification data. Here, we describe SUrface-protein Glycan And RNA-seq (SUGAR-seq); a method that enables detection and analysis of N-linked glycosylation, extracellular epitopes and the transcriptome at the single cell level. Integrated SUGAR-seq and glycoproteome analysis identified tumor infiltrating T cells with unique surface glycan properties that report their epigenetic and functional state. Overall design: Bulk 3' RNA sequencing of TILs from pooled day 14 B16-OVA tumors (CD8+, TCR+) which were FACS sorted on L-Pha low and L-Pha high.

多模态单细胞RNA测序（Multi-modal single cell RNA sequencing）能够精准绘制细胞分化状态的转录组与表型特征，但无法同时整合关键的翻译后修饰数据。
本文介绍了表面蛋白聚糖与RNA测序（SUrface-protein Glycan And RNA-seq，缩写为SUGAR-seq）技术：一种可在单细胞水平实现N-连接糖基化（N-linked glycosylation）、细胞外表位与转录组检测及分析的方法。整合SUGAR-seq与糖蛋白组学（glycoproteome）分析，可识别出具有独特表面聚糖特征的肿瘤浸润淋巴细胞（Tumor Infiltrating Lymphocytes，TILs），此类特征可反映其表观遗传与功能状态。
整体实验设计：对混合收集的第14天B16-OVA肿瘤来源的（CD8+、TCR+）肿瘤浸润淋巴细胞（TILs）进行批量3'端RNA测序，这些TILs此前通过荧光激活细胞分选（Fluorescence-Activated Cell Sorting，FACS）按照L-Pha低表达与L-Pha高表达表型进行了分选分群。