Visualising G-quadruplex DNA Dynamics in Live Cells by Fluorescence Lifetime Imaging Microscopy

Overview

Journal Nat Commun

Specialty Biology

Date 2021 Jan 9

PMID 33420085

Citations 54

Authors

Peter A Summers

Benjamin W Lewis

Jorge Gonzalez-Garcia

Rosa M Porreca

Aaron H M Lim

Paolo Cadinu

Nerea Martin-Pintado

David J Mann

Joshua B Edel

Jean Baptiste Vannier

Marina K Kuimova

Ramon Vilar

Affiliations

Soon will be listed here.

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.

Citing Articles

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