PUBLICATION
HI-NESS: a family of genetically encoded DNA labels based on a bacterial nucleoid-associated protein
- Authors
- Rashid, F.M., Mahlandt, E., van der Vaart, M., Boer, D.E.C., Varela Alvarez, M., Henneman, B., Brocken, D.J.W., Voskamp, P., Blok, A.J., Shimizu, T.S., Meijer, A.H., Luijsterburg, M.S., Goedhart, J., Crémazy, F.G.E., Dame, R.T.
- ID
- ZDB-PUB-211105-4
- Date
- 2021
- Source
- Nucleic acids research 50(2): e10 (Journal)
- Registered Authors
- Meijer, Annemarie H.
- Keywords
- none
- MeSH Terms
-
- Animals
- Bacterial Proteins/genetics
- Bacterial Proteins/metabolism*
- Biological Assay/methods*
- Cell Line
- Cloning, Molecular
- DNA Replication
- DNA, Bacterial/chemistry
- DNA, Bacterial/metabolism*
- DNA-Binding Proteins/genetics
- DNA-Binding Proteins/metabolism*
- Fluorescent Dyes
- Gene Expression
- Genetic Vectors
- Microscopy, Fluorescence
- Staining and Labeling/methods*
- PubMed
- 34734265 Full text @ Nucleic Acids Res.
Citation
Rashid, F.M., Mahlandt, E., van der Vaart, M., Boer, D.E.C., Varela Alvarez, M., Henneman, B., Brocken, D.J.W., Voskamp, P., Blok, A.J., Shimizu, T.S., Meijer, A.H., Luijsterburg, M.S., Goedhart, J., Crémazy, F.G.E., Dame, R.T. (2021) HI-NESS: a family of genetically encoded DNA labels based on a bacterial nucleoid-associated protein. Nucleic acids research. 50(2):e10.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping