Fluorescence-based melting assays for studying quadruplex ligands

Anne De Cian; Lionel Guittat; Markus Kaiser; Barbara Saccà; Samir Amrane; Anne Bourdoncle; Patrizia Alberti; Marie Paule Teulade-Fichou; Laurent Lacroix; Jean Louis Mergny

doi:10.1016/j.ymeth.2006.10.004

Fluorescence-based melting assays for studying quadruplex ligands

Anne De Cian
, Lionel Guittat
, Markus Kaiser
, Barbara Saccà
, Samir Amrane
, Anne Bourdoncle
, Patrizia Alberti
, Marie Paule Teulade-Fichou
, Laurent Lacroix
, Jean Louis Mergny

Research output: Contribution to journal › Article › peer-review

Abstract

The telomeric G-rich single-stranded DNA can adopt in vitro an intramolecular quadruplex structure, which has been shown to directly inhibit telomerase activity. The reactivation of this enzyme in immortalized and most cancer cells suggests that telomeres and telomerase are relevant targets in oncology, and telomere ligands and telomerase inhibitors have been proposed as new potential anticancer agents. In this paper, we have analysed the FRET method used to measure the stabilization and selectivity of quadruplex ligands towards the human telomeric G-quadruplex. The stabilization value depends on the nature of the fluorescent tags, the incubation buffer, and the method chosen for T_m calculation, complicating a direct comparison of the results obtained by different laboratories.

Original language	English
Pages (from-to)	183-195
Number of pages	13
Journal	Methods
Volume	42
Issue number	2
DOIs	https://doi.org/10.1016/j.ymeth.2006.10.004
Publication status	Published - 1 Jun 2007
Externally published	Yes

Keywords

DNA ligands
FRET
G-quadruplex
G-quartet
Telomerase inhibitor
Telomeres

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.ymeth.2006.10.004

Cite this

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title = "Fluorescence-based melting assays for studying quadruplex ligands",

abstract = "The telomeric G-rich single-stranded DNA can adopt in vitro an intramolecular quadruplex structure, which has been shown to directly inhibit telomerase activity. The reactivation of this enzyme in immortalized and most cancer cells suggests that telomeres and telomerase are relevant targets in oncology, and telomere ligands and telomerase inhibitors have been proposed as new potential anticancer agents. In this paper, we have analysed the FRET method used to measure the stabilization and selectivity of quadruplex ligands towards the human telomeric G-quadruplex. The stabilization value depends on the nature of the fluorescent tags, the incubation buffer, and the method chosen for Tm calculation, complicating a direct comparison of the results obtained by different laboratories.",

keywords = "DNA ligands, FRET, G-quadruplex, G-quartet, Telomerase inhibitor, Telomeres",

author = "\{De Cian\}, Anne and Lionel Guittat and Markus Kaiser and Barbara Sacc{\`a} and Samir Amrane and Anne Bourdoncle and Patrizia Alberti and Teulade-Fichou, \{Marie Paule\} and Laurent Lacroix and Mergny, \{Jean Louis\}",

note = "Funding Information: We thank Patrick Mailliet and Eliane Mandine (Sanofi-Aventis, Vitry, France) and Jean-Fran{\c c}ois Riou (Universit{\'e} de Reims) for helpful discussions. This work was supported in part by an ARC grant (No. 365), an EU FP6 “MolCancerMed” grant (LSHC-CT-2004-502943 to J.-L.M.) and a Fondation J{\'e}r{\^o}me Lejeune Ph.D. fellowship (to S.A.).",

year = "2007",

month = jun,

day = "1",

doi = "10.1016/j.ymeth.2006.10.004",

language = "English",

volume = "42",

pages = "183--195",

journal = "Methods",

issn = "1046-2023",

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}

TY - JOUR

T1 - Fluorescence-based melting assays for studying quadruplex ligands

AU - De Cian, Anne

AU - Guittat, Lionel

AU - Kaiser, Markus

AU - Saccà, Barbara

AU - Amrane, Samir

AU - Bourdoncle, Anne

AU - Alberti, Patrizia

AU - Teulade-Fichou, Marie Paule

AU - Lacroix, Laurent

AU - Mergny, Jean Louis

N1 - Funding Information: We thank Patrick Mailliet and Eliane Mandine (Sanofi-Aventis, Vitry, France) and Jean-François Riou (Université de Reims) for helpful discussions. This work was supported in part by an ARC grant (No. 365), an EU FP6 “MolCancerMed” grant (LSHC-CT-2004-502943 to J.-L.M.) and a Fondation Jérôme Lejeune Ph.D. fellowship (to S.A.).

PY - 2007/6/1

Y1 - 2007/6/1

N2 - The telomeric G-rich single-stranded DNA can adopt in vitro an intramolecular quadruplex structure, which has been shown to directly inhibit telomerase activity. The reactivation of this enzyme in immortalized and most cancer cells suggests that telomeres and telomerase are relevant targets in oncology, and telomere ligands and telomerase inhibitors have been proposed as new potential anticancer agents. In this paper, we have analysed the FRET method used to measure the stabilization and selectivity of quadruplex ligands towards the human telomeric G-quadruplex. The stabilization value depends on the nature of the fluorescent tags, the incubation buffer, and the method chosen for Tm calculation, complicating a direct comparison of the results obtained by different laboratories.

AB - The telomeric G-rich single-stranded DNA can adopt in vitro an intramolecular quadruplex structure, which has been shown to directly inhibit telomerase activity. The reactivation of this enzyme in immortalized and most cancer cells suggests that telomeres and telomerase are relevant targets in oncology, and telomere ligands and telomerase inhibitors have been proposed as new potential anticancer agents. In this paper, we have analysed the FRET method used to measure the stabilization and selectivity of quadruplex ligands towards the human telomeric G-quadruplex. The stabilization value depends on the nature of the fluorescent tags, the incubation buffer, and the method chosen for Tm calculation, complicating a direct comparison of the results obtained by different laboratories.

KW - DNA ligands

KW - FRET

KW - G-quadruplex

KW - G-quartet

KW - Telomerase inhibitor

KW - Telomeres

U2 - 10.1016/j.ymeth.2006.10.004

DO - 10.1016/j.ymeth.2006.10.004

M3 - Article

C2 - 17472900

AN - SCOPUS:34247641796

SN - 1046-2023

VL - 42

SP - 183

EP - 195

JO - Methods

JF - Methods

IS - 2

ER -

Fluorescence-based melting assays for studying quadruplex ligands

Abstract

Keywords

UN SDGs

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