ArticleScientific reports2018
A Novel Ultra-Stable, Monomeric Green Fluorescent Protein For Direct Volumetric Imaging of Whole Organs Using CLARITY.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 53 papers.
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Who cites it
53 citing papers in PubMed.
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- Stabilization versus flexibility: Detergent-dependent trade-offs in neurotensin receptor 1 GPCR ensembles.Protein science : a publication of the Protein Society · 2026Article
- Aggregation of Intrinsically Fluorescent Proteins: Combining Coarse-Grained Molecular Dynamics with Alphafold Predictions.ACS omega · 2025Article
- GroEL/ES chaperonin unfolds then encapsulates a nascent protein on the ribosome.Nature communications · 2025Article
- Stabilization versus flexibility: detergent-dependent trade-offs in neurotensin receptor 1 GPCR ensembles.bioRxiv : the preprint server for biology · 2025Article
- A multi-dentate, cooperative interaction between endo- and exo-ribonucleases within the bacterial RNA degradosome.Nucleic acids research · 2025Article
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- Stabilization of Condensate Interfaces Using Dynamic Protein Insertion.Journal of the American Chemical Society · 2025Article
- A highly stable monomeric red fluorescent protein for advanced microscopy.Nature methods · 2025Article
- Engineering Rubisco condensation in chloroplasts to manipulate plant photosynthesis.Plant biotechnology journal · 2025Article
- Accelerated enzyme engineering by machine-learning guided cell-free expression.Nature communications · 2025Article
- Long-Term Protein Synthesis with PURE in a Mesoscale Dialysis System.ACS synthetic biology · 2025Article
- In situ characterization of amine-forming enzymes shows altered oligomeric state.Protein science : a publication of the Protein Society · 2025Article
- Resolving chaperone-assisted protein folding on the ribosome at the peptide level.Nature structural & molecular biology · 2024Article
- Comparative studies of seafood and reptile α- and β-parvalbumins.Protein science : a publication of the Protein Society · 2024Article
- Autophagy adaptors mediate Parkin-dependent mitophagy by forming sheet-like liquid condensates.The EMBO journal · 2024Article
- De novo designed YK peptides forming reversible amyloid for synthetic protein condensates in mammalian cells.Nature communications · 2024Article
Corrections and comments
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Authors and funding
10 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recent advances in thick tissue clearing are enabling high resolution, volumetric fluorescence imaging of complex cellular networks. Fluorescent proteins (FPs) such as GFP, however, can be inactivated by the denaturing chemicals used to remove lipids in some tissue clearing methods. Here, we solved the crystal structure of a recently engineered ultra-stable GFP (usGFP) and propose that the two stabilising mutations, Q69L and N164Y, act to improve hydrophobic packing in the core of the protein and facilitate hydrogen bonding networks at the surface, respectively. usGFP was found to dimerise strongly, which is not desirable for some applications. A point mutation at the dimer interface, F223D, generated monomeric usGFP (muGFP). Neurons in whole mouse brains were virally transduced with either EGFP or muGFP and subjected to Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ hybridization-compatible Tissue-hYdrogel (CLARITY) clearing. muGFP fluorescence was retained after CLARITY whereas EGFP fluorescence was highly attenuated, thus demonstrating muGFP is a novel FP suitable for applications where high fluorescence stability and minimal self-association are required.
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