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Updated: Oct 21, 2025

Fluorescence Labeling to Visualize Low-Expressed Proteins in Zebrafish

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Published on: January 24, 2025

CRISPR/Cas9-Based Split Fluorescent Protein Tagging.

Gokul Kesavan¹, Anja Machate¹, Michael Brand¹

¹Center for Regenerative Therapies TU Dresden, Technische Universität Dresden, Dresden, Germany.

|September 8, 2021

Summary

This summary is machine-generated.

Genetically encoded fluorescent tags enable high-resolution live cell imaging. This study successfully integrates bright split fluorescent proteins in zebrafish using CRISPR/Cas9 for advanced protein labeling and live imaging applications.

Keywords:

CRISPR/Cas9 genetically encoded fluorescent protein tags knock-in split fluorescent protein

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Area of Science:

Cell Biology
Genetics
Biotechnology

Background:

Genetically encoded fluorescent tags like green fluorescent protein (GFP) have transformed cell biology by enabling live protein imaging.
Split fluorescent proteins (SFPs) offer a method to label proteins by inserting small fragments into their coding sequences.

Purpose of the Study:

To demonstrate the successful integration and utility of two novel split fluorescent proteins, mNeon green and sfCherry2, in zebrafish.
To validate the use of CRISPR/Cas9 gene editing for generating fluorescently tagged proteins in vivo.

Main Methods:

Integration of split fluorescent protein fragments (mNeon green and sfCherry2) into the zebrafish genome.
Utilizing CRISPR/Cas9 gene editing technology for targeted protein tagging.
Live imaging techniques, including time-lapse series, to assess protein localization and dynamics.

Main Results:

Successful integration of mNeon green and sfCherry2 split fluorescent proteins in zebrafish.
Demonstrated suitability of these tags for high signal-to-noise ratio live imaging, including time-lapse studies.
Validated CRISPR/Cas9 as an effective tool for in vivo generation of fluorescently tagged proteins.

Conclusions:

mNeon green and sfCherry2 are effective split fluorescent proteins for high-resolution live imaging in zebrafish.
CRISPR/Cas9 facilitates efficient generation of genetically encoded fluorescent protein tags in vivo.
These advancements provide powerful tools for studying protein dynamics and localization in live biological systems.