Identification of new DNA i-motif binding ligands through a fluorescent intercalator displacement assay† †Electronic supplementary information (ESI) available: Experimental, supporting fluorescence and UV-Vis data, Job plot, binding curves, CD spectra, melting temperatures and SPR data. See DOI: 10.1039/c7ob00710h

نویسندگان

  • Qiran Sheng
  • Joseph C Neaverson
  • Tasnim Mahmoud
  • Clare E M Stevenson
  • Susan E Matthews
  • Zoë A E Waller
چکیده

i-Motifs are quadruplex DNA structures formed from sequences rich in cytosine and held together by intercalated, hemi-protonated cytosine-cytosine base pairs. These sequences are prevalent in gene promoter regions and may play a role in gene transcription. Targeting these structures with ligands could provide a novel way to target genetic disease but there are very few ligands which have been shown to interact with i-motif DNA. Fluorescent intercalator displacement (FID) assays are a simple way to screen ligands against DNA secondary structures. Here we characterise how thiazole orange interacts with i-motif DNA and assess its ability for use in a FID assay. Additionally, we report FID-based ligand screening using thiazole orange against the i-motif forming sequence from the human telomere to reveal new i-motif binding compounds which have the potential for further development.

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منابع مشابه

Preferential targeting of i-motifs and G-quadruplexes by small molecules† †Electronic supplementary information (ESI) available: Experimental details, synthetic procedures, characterization data of compounds, 1H NMR and 13C NMR spectra, FRET melting, TO displacement, smFRET, lifetime data, CD spectra, western blot, dual luciferase, caspase assay. See DOI: 10.1039/c7sc02693e Click here for additional data file.

Department of Organic Chemistry, Indian A Jadavpur, Kolkata-700032, India. E-mail: oc Department of Physical Chemistry, Indian A Jadavpur, Kolkata-700032, India † Electronic supplementary information synthetic procedures, characterization d NMR spectra, FRET melting, TO displ spectra, western blot, dual luciferase, casp ‡ Present address: Department of Ch Education & Research, Bhopal, Bhopal-46...

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The importance of loop length on the stability of i-motif structures† †Electronic supplementary information (ESI) available: Experimental details, UV melting curves at 295 nm, and CD spectra at pH 5 and 8. See DOI: 10.1039/c4cc07279k Click here for additional data file.

Experimental All chemicals and oligonucleotides were purchased from Sigma-Aldrich. Oligonucleotides were purified by reverse phase column by Sigma-Aldrich and characteristic mass spectroscopy data is provided in table S1. UV spectroscopy was carried out using an Agilent Cary 100 with a temperature controlled 6 x 6 cell changer. CD spectra were collected on beamline B23 at Diamond Light Source. ...

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عنوان ژورنال:

دوره 15  شماره 

صفحات  -

تاریخ انتشار 2017