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Updated: Oct 22, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

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

Andrew Dunn¹, Yuqi Cai¹, Kentaro Iwasawa¹

¹Division of Gastroenterology, Hepatology & Nutrition, Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.

Stem Cell Reports

|August 27, 2021

Summary

This summary is machine-generated.

We developed POLYseq, a novel labeling system for cost-effective multiplexing in single-cell sequencing and live cell imaging. POLYseq enables efficient sample pooling and barcode delivery, overcoming current limitations in next-generation sequencing library preparation.

Keywords:

NGS cellular barcode iPS cell imaging organoid polymer

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Last Updated: Oct 22, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

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An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing

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Published on: May 23, 2018

Area of Science:

Biotechnology
Molecular Biology
Genomics

Background:

Next-generation sequencing (NGS) at the single-cell level is crucial for biological research.
Current NGS library preparation methods limit multiplexing, increasing costs due to separate library preparation for each sample.

Purpose of the Study:

To develop a novel, cost-effective labeling system for multiplexing in single-cell NGS and live cell imaging.
To enable rapid creation of custom sample pools for enhanced experimental scalability.

Main Methods:

Synthesis of a novel poly(β-amino) ester labeling system (POLYseq) using common reagents.
Delivery of fluorescent molecules or sample-distinguishing DNA barcodes via non-covalent binding.
Live imaging to track fluorescent POLYseq vectors and single-cell RNA sequencing to detect barcode labeling.

Main Results:

POLYseq efficiently delivers fluorescent molecules and DNA barcodes through non-covalent binding.
Cellular labeling propensity is determined by chemical formulation.
Lysosomal compartmentalization of POLYseq vectors was confirmed via live imaging.
Barcode labeling was detected in 90% of cells, enabling identification of mixed human and mouse cell lines.

Conclusions:

POLYseq offers a scalable and cost-effective solution for multiplexing in single-cell NGS.
The system demonstrates multifunctional applications in live cell imaging and sample pooling for genomics.
POLYseq overcomes limitations of current NGS library preparation, reducing costs and increasing efficiency.