ArticleScientific reports2017
CRISPR-Cas9 Mediated Labelling Allows for Single Molecule Imaging and Resolution.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
22 citing papers in PubMed, 47 citations in OpenAlex.
- Illuminating microtubule functions with small molecules: past, present and future of fluorescent tubulin-binding probes.RSC chemical biology · 2026Review
- Compression-dependent microtubule reinforcement enables cells to navigate confined environments.Nature cell biology · 2024Article
- ULK1 forms distinct oligomeric states and nanoscopic structures during autophagy initiation.Science advances · 2023Article
- Next-generation single virus tracking.Nature methods · 2022Article
- Superresolution microscopy localizes endogenous Dvl2 to Wnt signaling-responsive biomolecular condensates.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Post-translational polymodification of β1-tubulin regulates motor protein localisation in platelet production and function.Haematologica · 2022Article
- CRISPR/Cas9 Genome Editing vs. Over-Expression for Fluorescent Extracellular Vesicle-Labeling: A Quantitative Analysis.International journal of molecular sciences · 2021Article
- Development of a green reversibly photoswitchable variant of Eos fluorescent protein with fixation resistance.Molecular biology of the cell · 2021Article
- Article
- A versatile oblique plane microscope for large-scale and high-resolution imaging of subcellular dynamics.eLife · 2020Article
- Between life and death: strategies to reduce phototoxicity in super-resolution microscopy.Journal of physics D: Applied physics · 2020Review
- Topological data analysis quantifies biological nano-structure from single molecule localization microscopy.Bioinformatics (Oxford, England) · 2020Article
- Interactions at the cell membrane and pathways of internalization of nano-sized materials for nanomedicine.Beilstein journal of nanotechnology · 2020Review
- Optimised insert design for improved single-molecule imaging and quantification through CRISPR-Cas9 mediated knock-in.Scientific reports · 2019Article
- Review
- CRISPR-Cas9 a boon or bane: the bumpy road ahead to cancer therapeutics.Cancer cell international · 2019Review
- Measuring microtubule dynamics.Essays in biochemistry · 2018Review
- SRRF: Universal live-cell super-resolution microscopy.The international journal of biochemistry & cell biology · 2018Article
- Imaging platelet biogenesis in vivo.Research and practice in thrombosis and haemostasis · 2018Review
- Single-molecule fluorescence imaging: Generating insights into molecular interactions in virology.Journal of biosciences · 2018Review
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Single molecule imaging approaches like dSTORM and PALM resolve structures at 10-20 nm, and allow for unique insights into protein stoichiometry and spatial relationships. However, key obstacles remain in developing highly accurate quantitative single molecule approaches. The genomic tagging of PALM fluorophores through CRISPR-Cas9 offers an excellent opportunity for generating stable cell lines expressing a defined single molecule probe at endogenous levels, without the biological disruption and variability inherent to transfection. A fundamental question is whether these comparatively low levels of expression can successfully satisfy the stringent labelling demands of super-resolution SMLM. Here we apply CRISPR-Cas9 gene editing to tag a cytoskeletal protein (α-tubulin) and demonstrate a relationship between expression level and the subsequent quality of PALM imaging, and that spatial resolutions comparable to dSTORM can be achieved with CRISPR-PALM. Our approach shows a relationship between choice of tag and the total expression of labelled protein, which has important implications for the development of future PALM tags. CRISPR-PALM allows for nanoscopic spatial resolution and the unique quantitative benefits of single molecule localization microscopy through endogenous expression, as well as the capacity for super-resolved live cell imaging.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.