ArticleScientific reports2019
Optimised insert design for improved single-molecule imaging and quantification through CRISPR-Cas9 mediated knock-in.
Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
12 citing papers in PubMed.
- 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
- 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
- MYC amplifies gene expression through global changes in transcription factor dynamics.Cell reports · 2022Article
- Post-translational polymodification of β1-tubulin regulates motor protein localisation in platelet production and function.Haematologica · 2022Article
- Detection of genome-edited and endogenously expressed G protein-coupled receptors.The FEBS journal · 2021Review
- A nanoluciferase biosensor to investigate endogenous chemokine secretion and receptor binding.iScience · 2021Article
- Article
- Ligand-directed covalent labelling of a GPCR with a fluorescent tag in live cells.Communications biology · 2020Article
- Review
- CRISPR-Mediated Protein Tagging with Nanoluciferase to Investigate Native Chemokine Receptor Function and Conformational Changes.Cell chemical biology · 2020Article
- Multigene CRISPR/Cas9 genome editing of hybrid proline rich proteins (HyPRPs) for sustainable multi-stress tolerance in crops: the review of a promising approach.Physiology and molecular biology of plants : an international journal of functional plant biology · 2020Review
Corrections and comments
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Authors and funding
9 authors.
Funding
Abstract
The use of CRISPR-Cas9 genome editing to introduce endogenously expressed tags has the potential to address a number of the classical limitations of single molecule localisation microscopy. In this work we present the first systematic comparison of inserts introduced through CRISPR-knock in, with the aim of optimising this approach for single molecule imaging. We show that more highly monomeric and codon optimised variants of mEos result in improved expression at the TubA1B locus, despite the use of identical guides, homology templates, and selection strategies. We apply this approach to target the G protein-coupled receptor (GPCR) CXCR4 and show a further insert dependent effect on expression and protein function. Finally, we show that compared to over-expressed CXCR4, endogenously labelled samples allow for accurate single molecule quantification on ligand treatment. This suggests that despite the complications evident in CRISPR mediated labelling, the development of CRISPR-PALM has substantial quantitative benefits.
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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.