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Updated: May 3, 2026

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

Published on: April 14, 2015

Multiplexing clonality: combining RGB marking and genetic barcoding.

Kerstin Cornils¹, Lars Thielecke, Svenja Hüser

¹Research Department Cell and Gene Therapy, Department of Stem Cell Transplantation, University Medical Centre Hamburg-Eppendorf, Martinistrasse 52, Hamburg 20246, Germany, Institute for Medical Informatics and Biometry, Faculty of Medicine, Technische Universität Dresden, Dresden 01307, Germany, ALS Automated Lab Solutions GmbH, Jena 07747, Germany, Department of Neuropathology, Hannover Medical School, Institute of Pathology, Hannover 30625, Germany, Department of Internal Medicine I, University Medical Center Hamburg-Eppendorf, Hamburg 20246, Germany, Deep Sequencing Group SFB 655, Biotechnology Center, Technische Universität Dresden, Dresden 01307, Germany.

Nucleic Acids Research

|January 31, 2014

Summary

This summary is machine-generated.

This study introduces a novel RGB marking and DNA barcoding technique for precise clonal cell identification. This method enables unambiguous genetic marking of individual cells, aiding research in regeneration and disease.

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Multiplexed Single Cell mRNA Sequencing Analysis of Mouse Embryonic Cells

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Published on: January 7, 2020

Area of Science:

Biotechnology
Molecular Biology
Genetics

Background:

Clonal cell marking is crucial for understanding cell behavior and development.
Existing methods like RGB marking and DNA barcoding have limitations.
Combining these technologies offers enhanced capabilities for cell tracking.

Purpose of the Study:

To develop a novel technique integrating RGB marking with DNA barcoding for clonal cell identification.
To create and validate lentiviral vectors encoding fluorescent proteins and unique DNA barcodes.
To demonstrate the utility of this combined approach in cell line and primary cell studies, including disease modeling.

Main Methods:

Development of LeGO vectors encoding red, green, and blue fluorescent proteins with color-specific DNA barcodes.
Generation of complex plasmid libraries for producing barcoded lentiviral vector particles.
Application of single-cell polymerase chain reaction to decode barcode signatures in RGB-marked cells.

Main Results:

Successful RGB marking and DNA barcoding of cell lines and primary murine hepatocytes.
Proof-of-principle experiments confirmed clonal identity of cells with identical RGB colors.
Demonstrated application in studying clonal development of oncogene-induced leukemia in murine hematopoietic cells.

Conclusions:

The combined RGB marking and DNA barcoding technique provides unambiguous genetic marking of individual cells.
This novel method is applicable to studies of normal regeneration and malignant outgrowth.
Color-specific barcode signatures facilitate investigations into competitive repopulation dynamics under various experimental conditions.