title: Real-time and quantitative fluorescent live-cell imaging with quadruplex-specific red-edge probe (G4-REP).
abstract: The development of quadruplex-directed molecular diagnostic and therapy rely on mechanistic insights gained at both cellular and tissue levels by fluorescence imaging. This technique is based on fluorescent reporters that label cellular DNA and RNA quadruplexes to spatiotemporally address their complex cell biology. The photophysical characteristics of quadruplex probes usually dictate the modality of cell imaging by governing the selection of the light source (lamp, LED, laser), the optical light filters and the detection modality. Here, we report the characterizations of prototype from a new generation of quadruplex dye termed G4-REP (for quadruplex-specific red-edge probe) that provides fluorescence responses regardless of the excitation wavelength and modality (owing to the versatility gained through the red-edge effect), thus allowing for diverse applications and most imaging facilities. This is demonstrated by cell images (and associated quantifications) collected through confocal and multiphoton microscopy as well as through real-time live-cell imaging system over extended period, monitoring both non-cancerous and cancerous human cell lines. Our results promote a new way of designing versatile, efficient and convenient quadruplex-reporting dyes for tracking these higher-order nucleic acid structures in living human cells. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Daniela Montesarchio.
authors.affiliation: Faculty of Pharmaceutical Sciences, The University of British Columbia, Vancouver, Canada.
authors.forename: Sunny Y
authors.initials: SY
authors.lastname: Yang
authors.affiliation: Institut de Chimie Moléculaire, ICMUB CNRS UMR6302, UBFC Dijon, France.
authors.forename: Souheila
authors.initials: S
authors.lastname: Amor
authors.affiliation: Institut de Chimie Moléculaire, ICMUB CNRS UMR6302, UBFC Dijon, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Laguerre
authors.affiliation: Faculty of Pharmaceutical Sciences, The University of British Columbia, Vancouver, Canada. Electronic address: judy.wong@ubc.ca.
authors.forename: Judy M Y
authors.initials: JMY
authors.lastname: Wong
authors.affiliation: Institut de Chimie Moléculaire, ICMUB CNRS UMR6302, UBFC Dijon, France. Electronic address: david.monchaud@u-bourgogne.fr.
authors.forename: David
authors.initials: D
authors.lastname: Monchaud
mesh_terms: D001665:Binding Sites; D015374:Biosensing Techniques; D004273:DNA, Neoplasm / Q000737:chemistry / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D054856:G-Quadruplexes; D006151:Guanosine / Q000737:chemistry / Q000378:metabolism*; D057809:HEK293 Cells; D019073:HT29 Cells; D006801:Humans; D008024:Ligands; D018613:Microscopy, Confocal; D036641:Microscopy, Fluorescence, Multiphoton; D013329:Structure-Activity Relationship; D013997:Time Factors
chemical_list: D004273:DNA, Neoplasm; D005456:Fluorescent Dyes; D008024:Ligands; D006151:Guanosine
keywords: Cell imaging; Live-cell analysis system; Multiphoton microscopy; Red-edge effect; Smart quadruplex probe; Twice-as-smart quadruplex ligands