Despite evolving biological application of next-generation sequencing (NGS) at single-cell level, current techniques in NGS library preparation restrict multiplexing, necessitating the costly preparation of distinct libraries for each sample. Here, we report the development of a novel poly(β-amino) ester labeling system synthesized with inexpensive, common reagents, termed POLYseq, capable of efficiently delivering fluorescent molecules or sample-distinguishing DNA barcodes through non-covalent binding enabling rapid creation of custom sample pools. Chemical formulation was found to determine cellular labeling propensity. Live image-based tracking of fluorescent conjugated POLYseq vectors demonstrated lysosomal compartmentalization. Barcode labeling was uniformly detected across 90% of cells by single-cell RNA sequencing, allowing for the successful identification of human and mouse cultured cell lines from a single pool. These findings highlight the multifunctional applications of POLYseq in live cell imaging and NGS in a scalable and cost-effective manner.

尽管下一代测序 (NGS) 在单细胞水平上的生物学应用不断发展，但 NGS 文库制备中的当前技术限制了多路复用，因此需要为每个样品制备不同的文库，成本高昂。在这里，我们报告了一种新型聚（β-氨基）酯标记系统的开发，该系统由廉价的常用试剂​​合成，称为 POLYseq，能够通过非共价结合有效地传递荧光分子或区分样本的 DNA 条形码，从而能够快速创建定制样本池。发现化学制剂可确定细胞标记倾向。基于实时图像的荧光共轭 POLYseq 载体跟踪证明了溶酶体区室化。通过单细胞 RNA 测序在 90% 的细胞中均一地检测到条形码标记，允许从单个池中成功识别人和小鼠培养的细胞系。这些发现突出了 POLYseq 在活细胞成像和 NGS 中以可扩展且具有成本效益的方式的多功能应用。