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Updated: Mar 29, 2026
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Assessing Protein Interactions in Live-Cells with FRET-Sensitized Emission
Published on: April 22, 2021
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Intensity modulation of trichromatic split fluorescent proteins for live cell mapping.
Mamoru Ishii1, Tomoaki Kinjo2, Yohei Kondo3
1Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan; Institute of Industrial Science, The University of Tokyo, Tokyo, Japan.
Cell Reports Methods
|March 27, 2026
Summary
Researchers developed Caterpie, a novel cell labeling system using engineered split fluorescent proteins. This technology allows for accurate identification of 20 cell populations, overcoming limitations of current multiplexed cell labeling methods.
Area of Science:
- Cell biology
- Biotechnology
- Molecular imaging
Background:
- Multiplexed cell labeling is crucial for understanding complex biological systems.
- Current fluorescent protein techniques face challenges with limited discrimination and large gene sizes.
- Stochastic color selection and tandem repeats hinder precise cell population identification.
Purpose of the Study:
- To develop a novel cell labeling system with enhanced discrimination power and reduced gene size.
- To engineer split fluorescent proteins for deterministic and high-fidelity cell population identification.
- To overcome limitations of existing multiplexed cell labeling methods.
Main Methods:
- Development of Caterpie, a rationally designed system utilizing engineered split fluorescent proteins.
- Computational structure-guided design of enhanced split mNeonGreen3A and split sfCherry3C variants.
- Creation of a systematic library of trichromatic 11th β-strand tags with up to 12 tandem repeats.
Main Results:
- Deterministic identification of 20 distinct cell populations with 97% accuracy.
- Engineered split fluorescent proteins achieved performance comparable to split CFP2.
- Reduced gene sizes compared to traditional tandem repeat fluorescent proteins.
- Predictable, high-fidelity labeling for precise cell targeting.
Conclusions:
- Caterpie system significantly improves multiplexed cell labeling capabilities.
- The technology enables precise targeting and simultaneous identification of multiple defined cell populations.
- Engineered split fluorescent proteins offer a powerful tool for advanced cell biology research.
