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Updated: Oct 21, 2025
09:38
Fluorescence Labeling to Visualize Low-Expressed Proteins in Zebrafish
Published on: January 24, 2025
927
CRISPR/Cas9-Based Split Fluorescent Protein Tagging.
Gokul Kesavan1, Anja Machate1, Michael Brand1
1Center for Regenerative Therapies TU Dresden, Technische Universität Dresden, Dresden, Germany.
Zebrafish
|September 8, 2021
Summary
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.
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.
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