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Related Experiment Video

Updated: Dec 29, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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High-complexity extracellular barcoding using a viral hemagglutinin.

Gavriel Mullokandov1, Gayathri Vijayakumar2, Paul Leon2

  • 1Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029.

Proceedings of the National Academy of Sciences of the United States of America
|January 29, 2020
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel protein barcoding system using influenza hemagglutinins to track cellular clones. This method overcomes limitations of fluorescent proteins and DNA barcodes for pooled screens and lineage tracing in vivo.

Keywords:
cell barcodingprotein engineeringvirology

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Area of Science:

  • Molecular Biology
  • Genetics
  • Immunology

Background:

  • Single-cell sequencing reveals tissue heterogeneity.
  • Resolving cellular clones is crucial for pooled screens and lineage tracing.
  • Existing methods like fluorescent proteins and DNA barcodes have limitations.

Purpose of the Study:

  • To engineer a genetically encoded cell-surface protein barcoding system.
  • To overcome limitations of current barcoding technologies.
  • To enable clonal tracking in vivo.

Main Methods:

  • Engineered influenza virus hemagglutinins for cell-surface protein barcoding.
  • Utilized antibodies paired to hemagglutinins with escape mutations.
  • Developed an exponential protein barcoding system.

Main Results:

  • Successfully created a protein-level cell barcode system.
  • Enabled labeling of 128 distinct clones using seven antibodies.
  • Demonstrated proof of principle for in vivo clonal tracking.

Conclusions:

  • The engineered protein barcoding system offers a scalable solution for clonal analysis.
  • This technology overcomes limitations of fluorescent proteins and DNA barcodes.
  • Provides a foundation for advanced clonal lineage tracing in complex biological systems.