POLYseq: A poly(ß-amino ester)-based vector for multifunctional cellular barcoding

Despite evolving stem cell and organoid application of next-generation sequencing (NGS) at single cell level, current techniques in NGS library preparation are restrictive as individual samples within a single library are indistinguishable, necessitating the laborious and costly preparation of distinct libraries for each sample. To combat this challenge, we report the development of a novel poly(ß-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 libraries. Delivery of Next-Gen readable ssDNA barcodes by POLYseq into individual human liver organoid samples and pooled cell lines