Ligands playing musical chairs with G-quadruplex DNA: a rapid and simple displacement assay for identifying selective G-quadruplex binders.
We report here the details of G4-FID (G-quadruplex fluorescent intercalator displacement), a simple method aiming at evaluating quadruplex-DNA binding affinity and quadruplex- over duplex-DNA selectivity of putative ligands. This assay is based on the loss of fluorescence upon displacement of thiazole orange from quadruplex- and duplex-DNA matrices. The original protocol was tested using various quadruplex- and duplex-DNA targets, and with a wide panel of G-quadruplex ligands belonging to different families (i.e. from quinacridines to metallo-organic ligands) likely to display various binding modes. The reliability of the assay is further supported by comparisons with FRET-melting and ESI-MS assays.
Monchaud
D
D
Institut Curie, Section Recherche, CNRS UMR176, Centre Universitaire Paris XI, Bat. 110, 91405 Orsay, France.
Allain
C
C
Bertrand
H
H
Smargiasso
N
N
Rosu
F
F
Gabelica
V
V
De Cian
A
A
Mergny
J-L
JL
Teulade-Fichou
M-P
MP
D000166:Acridines / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D052160:Benzothiazoles / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D054856:G-Quadruplexes; D008024:Ligands; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009942:Organometallic Compounds / Q000378:metabolism; D011804:Quinolines / Q000378:metabolism; D011806:Quinolinium Compounds / Q000378:metabolism; D012492:Salts / Q000494:pharmacology; D012680:Sensitivity and Specificity; D013997:Time Factors
D000166:Acridines; D052160:Benzothiazoles; D008024:Ligands; D009841:Oligonucleotides; D009942:Organometallic Compounds; D011804:Quinolines; D011806:Quinolinium Compounds; D012492:Salts; C051000:thiazole orange; D004247:DNA