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Visualising G-quadruplex DNA dynamics in live cells by fluorescence lifetime imaging microscopy

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Title: Visualising G-quadruplex DNA dynamics in live cells by fluorescence lifetime imaging microscopy
Authors: Vilar Compte, R
Summers, P
Lewis, B
Gonzalez-Garcia, J
Porreca, RM
Lim, A
Cadinu, P
Martin-Pintado, N
Mann, D
Edel, J
Vannier, JB
Kuimova, M
Vilar Compte, R
Item Type: Journal Article
Abstract: Guanine rich regions of oligonucleotides fold into quadruple-stranded structures called G-quadruplexes (G4s). Increasing evidence suggests that these G4 structures form in vivo and play a crucial role in cellular processes. However, their direct observation in live cells remains a challenge. Here we demonstrate that a fluorescent probe (DAOTA-M2) in conjunction with fluorescence lifetime imaging microscopy (FLIM) can identify G4s within nuclei of live and fixed cells. We present a FLIM-based cellular assay to study the interaction of non-fluorescent small molecules with G4s and apply it to a wide range of drug candidates. We also demonstrate that DAOTA-M2 can be used to study G4 stability in live cells. Reduction of FancJ and RTEL1 expression in mammalian cells increases the DAOTA-M2 lifetime and therefore suggests an increased number of G4s in these cells, implying that FancJ and RTEL1 play a role in resolving G4 structures in cellulo.
Date of Acceptance: 27-Nov-2020
URI: http://hdl.handle.net/10044/1/85968
DOI: 10.1038/s41467-020-20414-7
ISSN: 2041-1723
Publisher: Nature Research
Journal / Book Title: Nature Communications
Volume: 12
Issue: 162
Copyright Statement: © The Author(s) 2021
Sponsor/Funder: Biotechnology and Biological Sciences Research Council (BBSRC)
Commission of the European Communities
Funder's Grant Number: BB/P026109/1
637798
Keywords: Animals
Cell Line, Tumor
DNA
DNA Helicases
Fanconi Anemia Complementation Group Proteins
Fibroblasts
Fluorescent Dyes
G-Quadruplexes
Gene Knockdown Techniques
Humans
Indoles
Intravital Microscopy
Mice
Microscopy, Fluorescence
Molecular Imaging
RNA Helicases
Cell Line, Tumor
Fibroblasts
Animals
Humans
Mice
Indoles
DNA Helicases
RNA Helicases
DNA
Fluorescent Dyes
Microscopy, Fluorescence
Fanconi Anemia Complementation Group Proteins
G-Quadruplexes
Gene Knockdown Techniques
Molecular Imaging
Intravital Microscopy
Publication Status: Published
Online Publication Date: 2021-01-08
Appears in Collections:Chemistry
Biological and Biophysical Chemistry
Institute of Clinical Sciences
Faculty of Natural Sciences



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