ArticleCell reports methods2026
Intensity modulation of trichromatic split fluorescent proteins for live cell mapping.
Article in Cell reports methods, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Current fluorescent protein-based multiplexed cell labeling techniques suffer from limited discrimination power due to stochastic color selection and large gene sizes from tandem repeats of multiple fluorescent proteins. We developed Caterpie, a rationally designed system using engineered split fluorescent proteins that enables deterministic identification of 20 distinct cell populations with 97% accuracy and reduced gene sizes. Through computational structure-guided design, we engineered enhanced split mNeonGreen3A and split sfCherry3C variants that achieve performance comparable to split CFP2, the best-performing split fluorescent protein. Our systematic library of trichromatic 11th β-strand tags with up to 12 tandem repeats enables predictable, high-fidelity labeling for precise cell targeting. This technology addresses critical limitations in simultaneous identification of multiple defined cell populations.
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