ArticleGenetics2021
Split-wrmScarlet and split-sfGFP: tools for faster, easier fluorescent labeling of endogenous proteins in Caenorhabditis elegans.
Article in Genetics, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers.
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44 citing papers in PubMed.
- Recent developments of fluorescent proteins and biosensors driven by cross-disciplinary insights.Nature methods · 2026Review
- Systematic identification of germ granule proteins reveals specialized roles in RNAi and small RNA inheritance.Genes & development · 2026Article
- Functions and trafficking mechanisms of RIC-8 inMolecular biology of the cell · 2026Article
- Mechanosensory encoding of surface mechanics optimizes locomotion.Nature communications · 2026Article
- A genetically encoded sensor of ionic stress links cellular proton dynamics to sleep.Science advances · 2026Article
- Efficient Endogenous Tagging in the Sea Urchin,bioRxiv : the preprint server for biology · 2026Article
- Endogenous autophagosome reporters reveal redundant and non-redundant roles of LGG-1 and LGG-2 inbioRxiv : the preprint server for biology · 2026Article
- A conserved Arf-GEF modulates axonal integrity through RAB-35 by altering neuron-epidermal attachment.Journal of cell science · 2026Article
- Intensity modulation of trichromatic split fluorescent proteins for live cell mapping.Cell reports methods · 2026Article
- PES-8 is required for Cytoskeletal Organization and Contractility in thebioRxiv : the preprint server for biology · 2026Article
- ER discontinuities are common inbioRxiv : the preprint server for biology · 2026Article
- An apical junction protein antagonizes mechanosensitive calcium signaling to establish stochastic choices of olfactory neuron subtypes.bioRxiv : the preprint server for biology · 2026Article
- Cell-specific roles for the conserved Galpha chaperone RIC-8 in cilia biology.bioRxiv : the preprint server for biology · 2026Article
- A genetically encoded ionic-stress sensor reveals protons as a sleep driver.bioRxiv : the preprint server for biology · 2026Article
- Systematic Identification of Germ Granule Proteins Reveals Specialized Roles in RNAi and Small RNA Inheritance.bioRxiv : the preprint server for biology · 2026Article
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- TurboID-based proximity labeling identifies novel germline proteins that maintain E granule integrity and small RNA homeostasis in C. elegans.Science China. Life sciences · 2025Article
- A single-copy knock-in system: one plasmid to target all chromosomes in C. elegans.G3 (Bethesda, Md.) · 2025Article
- Bipartite Genetically Encoded Biosensors to Sense Calcium Ion Dynamics at Membrane-Membrane Contact Sites.Analytical chemistry · 2025Article
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9 authors.
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Abstract
We create and share a new red fluorophore, along with a set of strains, reagents and protocols, to make it faster and easier to label endogenous Caenorhabditis elegans proteins with fluorescent tags. CRISPR-mediated fluorescent labeling of C. elegans proteins is an invaluable tool, but it is much more difficult to insert fluorophore-size DNA segments than it is to make small gene edits. In principle, high-affinity asymmetrically split fluorescent proteins solve this problem in C. elegans: the small fragment can quickly and easily be fused to almost any protein of interest, and can be detected wherever the large fragment is expressed and complemented. However, there is currently only one available strain stably expressing the large fragment of a split fluorescent protein, restricting this solution to a single tissue (the germline) in the highly autofluorescent green channel. No available C. elegans lines express unbound large fragments of split red fluorescent proteins, and even state-of-the-art split red fluorescent proteins are dim compared to the canonical split-sfGFP protein. In this study, we engineer a bright, high-affinity new split red fluorophore, split-wrmScarlet. We generate transgenic C. elegans lines to allow easy single-color labeling in muscle or germline cells and dual-color labeling in somatic cells. We also describe a novel expression strategy for the germline, where traditional expression strategies struggle. We validate these strains by targeting split-wrmScarlet to several genes whose products label distinct organelles, and we provide a protocol for easy, cloning-free CRISPR/Cas9 editing. As the collection of split-FP strains for labeling in different tissues or organelles expands, we will post updates at doi.org/10.5281/zenodo.3993663.
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