Single‐Cell Glycan and RNA Sequencing (scGR‐seq)

Single-cell multi-omics analysis has emerged as a crucial tool in modern biology due to its capacity to uncover the intricate molecular heterogeneity within individual cells. Glycan structural diversity on the cell surface creates a "cell signature" that varies by cell type and state. However, single-cell glycan analysis remains a challenge. This protocol presents a series of techniques and procedures that enable the simultaneous measurements of glycan and RNA at the single-cell level via DNA-barcoded lectin-based sequencing (scGR-seq). The techniques include (1) the preparation of DNA-barcoded lectins, (2) a step-by-step protocol for single-cell glycan/RNA sequencing, and (3) a data analysis approach for the integration of glycan and RNA data. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Preparation of DNA-barcoded lectins Basic Protocol 2: Single-cell glycan/RNA sequencing.