ArticlePLoS biology2024
AlphaFold2-guided engineering of split-GFP technology enables labeling of endogenous tubulins across species while preserving function.
Article in PLoS biology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- Gain-of-function suppressors restore microtubule dynamics and rescue dominant-negative tubulinopathies.Nature cell biology · 2026Article
- Towards a systems-level view of the microtubule cytoskeleton and its functions in physiology and disease.Nature reviews. Molecular cell biology · 2026Review
- Screening the composition of tubulin isotypes reveals the most abundant TUBB4B for ciliary polarity in ependymal cells.Journal of molecular cell biology · 2026Article
- TaggingbioRxiv : the preprint server for biology · 2026Article
- WormTagDB: a systematic survey of endogenously tagged proteins in Caenorhabditis elegans and roadmap toward the tagged proteome.G3 (Bethesda, Md.) · 2026Article
- Illuminating microtubule functions with small molecules: past, present and future of fluorescent tubulin-binding probes.RSC chemical biology · 2026Review
- Technological advances in visualizing and rewiring microtubules during plant development.Journal of experimental botany · 2025Review
- WormTagDB: A Systematic Survey of Endogenously Tagged Proteins inbioRxiv : the preprint server for biology · 2025Article
- Evolution of split-protein technologies in virology: From mechanistic discovery and diagnostics to therapeutic promise.Journal of virus eradication · 2025Review
- Kinesins inOpen biology · 2025Review
- Functional fluorescence labeling of actins in live-cell system using engineered split-GFP technology.Molecular biology of the cell · 2025Article
- A tubulin-MAPKKK pathway engages tubulin isotype interaction for neuroprotection.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Fluorescent protein tags for human tropomyosin isoform comparison.Biology open · 2025Article
- Tissue-specific consequences of tag fusions on protein expression in transgenic mice.PLoS genetics · 2025Article
- Alpha-tubulin tails regulate axoneme differentiation.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Peri-centrosomal localization of small interfering RNAs in C. elegans.Science China. Life sciences · 2025Article
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11 authors.
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Abstract
Dynamic properties are essential for microtubule (MT) physiology. Current techniques for in vivo imaging of MTs present intrinsic limitations in elucidating the isotype-specific nuances of tubulins, which contribute to their versatile functions. Harnessing the power of the AlphaFold2 pipeline, we engineered a strategy for the minimally invasive fluorescence labeling of endogenous tubulin isotypes or those harboring missense mutations. We demonstrated that a specifically designed 16-amino acid linker, coupled with sfGFP11 from the split-sfGFP system and integration into the H1-S2 loop of tubulin, facilitated tubulin labeling without compromising MT dynamics, embryonic development, or ciliogenesis in Caenorhabditis elegans. Extending this technique to human cells and murine oocytes, we visualized MTs with the minimal background fluorescence and a pathogenic tubulin isoform with fidelity. The utility of our approach across biological contexts and species set an additional paradigm for studying tubulin dynamics and functional specificity, with implications for understanding tubulin-related diseases known as tubulinopathies.
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