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

Stem Cell Reports. 2021 Sep 14;16(9):2149-2158. doi: 10.1016/j.stemcr.2021.07.020. Epub 2021 Aug 26.


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.

Keywords: NGS; cellular barcode; iPS cell; imaging; organoid; polymer.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Cell Differentiation / genetics
  • DNA Barcoding, Taxonomic / methods*
  • Flow Cytometry
  • Fluorescent Dyes
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Organoids
  • Polymers / chemistry*
  • Single-Cell Analysis / methods
  • Staining and Labeling / methods*


  • Fluorescent Dyes
  • Polymers
  • poly(beta-amino ester)