TY - JOUR
T1 - HI-NESS
T2 - a family of genetically encoded DNA labels based on a bacterial nucleoid-associated protein
AU - Rashid, Fatema-Zahra M.
AU - Mahlandt, Eike
AU - van der Vaart, Michiel
AU - Boer, Daphne E.C.
AU - Alvarez, Monica Varela
AU - Henneman, Bram
AU - Brocken, Daan J.W.
AU - Voskamp, Patrick
AU - Blok, Anneloes J.
AU - Shimizu, Thomas S.
AU - Meijer, Annemarie H.
AU - Luijsterburg, Martijn S.
AU - Goedhart, Joachim
AU - Crémazy, Frédéric G.E.
AU - Dame, Remus T.
PY - 2022/1/25
Y1 - 2022/1/25
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85125017417&partnerID=8YFLogxK
U2 - 10.1093/nar/gkab993
DO - 10.1093/nar/gkab993
M3 - Article
SN - 0305-1048
VL - 50
JO - Nucleic acids research
JF - Nucleic acids research
IS - 2
ER -