HI-NESS: a family of genetically encoded DNA labels based on a bacterial nucleoid-associated protein
| Autor: | Patrick Voskamp, Daphne E. C. Boer, Monica Varela Alvarez, Martijn S. Luijsterburg, Fatema-Zahra M Rashid, Frédéric Crémazy, Daan J.W. Brocken, Thomas S Shimizu, Joachim Goedhart, Remus T. Dame, Annemarie H. Meijer, Michiel van der Vaart, Eike K. Mahlandt, Anneloes Blok, Bram Henneman |
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| Přispěvatelé: | Molecular Cytology (SILS, FNWI) |
| Jazyk: | angličtina |
| Rok vydání: | 2022 |
| Předmět: |
DNA Replication
DNA Bacterial AcademicSubjects/SCI00010 DNA damage Genetic Vectors Gene Expression Biology Genome Cell Line chemistry.chemical_compound Narese/3 Bacterial Proteins Transcription (biology) Genetics Animals Cloning Molecular Fluorescent Dyes Staining and Labeling Chromosome Bacterial nucleoid Cell biology Chromatin DNA-Binding Proteins Microscopy Fluorescence chemistry Nucleic acid Methods Online Biological Assay DNA |
| Zdroj: | Nucleic Acids Research, 50(2). OXFORD UNIV PRESS Nucleic Acids Research, 50(2). Oxford University Press (OUP) Nucleic Acids Research, 50(2) Nucleic Acids Research, 50(2):e10. Oxford University Press Nucleic Acids Research |
| ISSN: | 0305-1048 |
| Popis: | The interplay between three-dimensional chromosome organisation and genomic processes such as replication and transcription necessitates in vivo studies of chromosome dynamics. Fluorescent organic dyes are often used for chromosome labelling in vivo. The mode of binding of these dyes to DNA cause its distortion, elongation, and partial unwinding. The structural changes induce DNA damage and interfere with the binding dynamics of chromatin-associated proteins, consequently perturbing gene expression, genome replication, and cell cycle progression. We have developed a minimally-perturbing, genetically encoded fluorescent DNA label consisting of a (photo-switchable) fluorescent protein fused to the DNA-binding domain of H-NS — a bacterial nucleoid-associated protein. We show that this DNA label, abbreviated as HI-NESS (H-NS-based indicator for nucleic acid stainings), is minimally-perturbing to genomic processes and labels chromosomes in eukaryotic cells in culture, and in zebrafish embryos with preferential binding to AT-rich chromatin. |
| Databáze: | OpenAIRE |
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