Isolation of G-quadruplex DNA using NMM-sepharose affinity chromatography.
DNA can adopt a variety of non-standard conformations, including structures known as G-quadruplexes (G4-DNA), which consist of stacked tetrads of guanines. There are growing indications that G4-DNA is of biological importance, including evidence that it plays roles in telomere function, DNA recombination and the regulation of transcription and translation. However, it has been difficult to obtain direct, physical evidence for the presence of G-quadruplex DNA in vivo due, in part, to a lack of tools for G4-DNA identification. Here, we describe a method for coupling the G4-DNA binding ligand N-methyl mesoporphyrin IX (NMM) to a Sepharose resin, and demonstrate the ability of the resin to bind tightly and selectively to DNA oligonucleotides with the capacity to form G4-DNA. This technique might also be extended to examine genomic distributions of G4-DNA isolated from in vivo sources.
Smith
Jasmine S
JS
Department of Pathology, Cancer Biology Program, and Institute on Aging, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
Johnson
F Brad
FB
D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D054856:G-Quadruplexes; D006147:Guanine / Q000737:chemistry; D006801:Humans; D008652:Mesoporphyrins / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry; D016615:Telomere / Q000737:chemistry*
D008652:Mesoporphyrins; C031468:mesoporphyrin IX; D006147:Guanine; D004247:DNA; D012685:Sepharose