Fluorescence-based melting assays for studying quadruplex ligands.
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.
De Cian
Anne
A
Laboratoire de Biophysique, Muséum National d'Histoire Naturelle, USM 503, INSERM UR 565, CNRS UMR 5153, 43 rue Cuvier, 75231 Paris Cedex 05, France.
Guittat
Lionel
L
Kaiser
Markus
M
Saccà
Barbara
B
Amrane
Samir
S
Bourdoncle
Anne
A
Alberti
Patrizia
P
Teulade-Fichou
Marie-Paule
MP
Lacroix
Laurent
L
Mergny
Jean-Louis
JL
D001483:Base Sequence; D002021:Buffers; D004247:DNA; D004277:DNA, Single-Stranded / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer; D054856:G-Quadruplexes; D006147:Guanine / Q000737:chemistry; D006358:Hot Temperature; D006801:Humans; D008024:Ligands; D008956:Models, Chemical; D009690:Nucleic Acid Conformation; D019098:Telomerase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D016615:Telomere / Q000737:chemistry*
D002021:Buffers; D004277:DNA, Single-Stranded; D004791:Enzyme Inhibitors; D008024:Ligands; D006147:Guanine; D004247:DNA; D019098:Telomerase