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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
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Related Experiment Video

Updated: Sep 15, 2025

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications
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Published on: April 14, 2015

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Cell barcoding with tandem fluorescent proteins enables high-throughput signaling dynamics analysis.

Jhen-Wei Wu, Jr-Ming Yang, Suyang Wang

    Biorxiv : the Preprint Server for Biology
    |July 16, 2025
    PubMed
    Summary

    We developed single chain tandem fluorescent protein (sctFP) barcodes for cell barcoding. These barcodes offer high diversity and simple readout for applications like lineage tracing and genetic screening.

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    Area of Science:

    • Biotechnology
    • Molecular Biology
    • Cell Biology

    Background:

    • Cell barcodes are crucial for various biological experiments, including lineage tracing and single-cell analysis.
    • Existing methods lack high diversity, live-cell compatibility, and simplified readout.
    • There is a need for improved barcoding strategies to enhance throughput and scalability.

    Purpose of the Study:

    • To introduce and characterize single chain tandem fluorescent protein (sctFP) barcodes.
    • To demonstrate the utility of sctFP barcodes for multiplexed biosensing and cell tracking.
    • To provide a robust and scalable cell barcoding solution for diverse applications.

    Main Methods:

    • Construction of sctFP barcodes by linking different fluorescent proteins (FPs) in single polypeptide chains.
    • Utilizing fluorescence signal intensity ratios at different wavelengths for sctFP differentiation.
    • Application of sctFP barcodes with genetically encoded fluorescent biosensors.

    Main Results:

    • sctFP barcodes generated from cnidarian FPs showed reliable differentiation based on spectral signal ratios.
    • Prokaryotic FPs were less suitable due to cofactor requirements.
    • Demonstrated multiplexed tracking of signaling activities using sctFP-compatible biosensors.
    • Enabled simplified imaging and analysis pipelines for high-throughput applications.

    Conclusions:

    • sctFP barcodes offer a robust, scalable, and versatile method for cell barcoding.
    • This strategy enhances multiplexing capabilities for fluorescent biosensors.
    • sctFP barcodes are compatible with a wide range of biosensor types and experimental contexts.