Development of microfluidic platforms for the synthesis of metal complexes and evaluation of their DNA affinity using online FRET melting assays

Rakers, Viktoria; Cadinu, Paolo; Edel, Joshua B.; Vilar, Ramón

doi:10.1039/c8sc00528a

Cited by 18 publications

(13 citation statements)

References 52 publications

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“…Rakers et al developed a novel microfluidic platform to synthesize novel G4 binders and evaluate their affinity and using a real-time online FRET assay [ 200 ]. Furthermore, their strategy enabled the high-throughput online analysis of several G4 binders.…”

Section: Methods To Characterize G4/ligand Interactionsmentioning

confidence: 99%

G-Quadruplexes and Their Ligands: Biophysical Methods to Unravel G-Quadruplex/Ligand Interactions

et al. 2021

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Progress in the design of G-quadruplex (G4) binding ligands relies on the availability of approaches that assess the binding mode and nature of the interactions between G4 forming sequences and their putative ligands. The experimental approaches used to characterize G4/ligand interactions can be categorized into structure-based methods (circular dichroism (CD), nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography), affinity and apparent affinity-based methods (surface plasmon resonance (SPR), isothermal titration calorimetry (ITC) and mass spectrometry (MS)), and high-throughput methods (fluorescence resonance energy transfer (FRET)-melting, G4-fluorescent intercalator displacement assay (G4-FID), affinity chromatography and microarrays. Each method has unique advantages and drawbacks, which makes it essential to select the ideal strategies for the biological question being addressed. The structural- and affinity and apparent affinity-based methods are in several cases complex and/or time-consuming and can be combined with fast and cheap high-throughput approaches to improve the design and development of new potential G4 ligands. In recent years, the joint use of these techniques permitted the discovery of a huge number of G4 ligands investigated for diagnostic and therapeutic purposes. Overall, this review article highlights in detail the most commonly used approaches to characterize the G4/ligand interactions, as well as the applications and types of information that can be obtained from the use of each technique.

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Section: Methods To Characterize G4/ligand Interactionsmentioning

confidence: 99%

G-Quadruplexes and Their Ligands: Biophysical Methods to Unravel G-Quadruplex/Ligand Interactions

et al. 2021

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“…N , N′ ‐Bis[4‐[[2‐(trimethylammonio)ethyl]oxy]salicylidene]‐ o ‐phenyldiamine dibromide (3) : 4‐(4‐Formyl‐3‐hydroxyphenoxy)‐ N , N , N ,‐trimethylethan‐1‐ammonium bromide ( 4 , synthesized as previously reported) (800 mg, 2.63 mmol, 2 equiv) was dissolved in 50 mL absolute ethanol. 1,2‐Phenylenediamine (142.1 mg, 1.31 mmol, 1 equiv) was added and the reaction refluxed for 5 hours under an inert atmosphere.…”

Section: Methodsmentioning

confidence: 99%

An Octahedral Cobalt(III) Complex with Axial NH₃ Ligands that Templates and Selectively Stabilises G‐quadruplex DNA

Ruehl

Lim

Kench

et al. 2019

Chemistry A European J

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Guanine‐rich sequences of DNA are known to readily fold into tetra‐stranded helical structures known as G‐quadruplexes (G4). Due to their biological relevance, G4s are potential anticancer drug targets and therefore there is significant interest in molecules with high affinity for these structures. Most G4 binders are polyaromatic planar compounds which π–π stack on the G4′s guanine tetrad. However, many of these compounds are not very selective since they can also intercalate into duplex DNA. Herein we report a new class of binder based on an octahedral cobalt(III) complex that binds to G4 via a different mode involving hydrogen bonding, electrostatic interactions and π–π stacking. We show that this new compound binds selectivity to G4 over duplex DNA (particularly to the G‐rich sequence of the c‐myc promoter). This new octahedral complex also has the ability to template the formation of G4 DNA from the unfolded sequence. Finally, we show that upon binding to G4, the complex prevents helicase Pif1‐p from unfolding the c‐myc G4 structure.

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“…[14,15] An example of this, are metal salphen ands alen complexes, which have been extensively studied over the past 10 years (see Figure 1). [16,17] Aw ide range of derivatives with different metals (and hence geometries), [18][19][20] salphen/salen core structure [21,22] and nature/number/position of substituents [19,[23][24][25][26][27] have been reporteda se xcellent G4 DNA binders. In spite of this, there is still little information regardingt he selectivity of metal-salphens against several different G4 DNA structures.…”

Section: Introductionmentioning

confidence: 99%

Binding Studies of Metal–Salphen and Metal–Bipyridine Complexes towards G‐Quadruplex DNA

Łęczkowska

González‐García

Pérez‐Arnaiz

et al. 2018

Chemistry A European J

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The proposed in vivo formation of G-quadruplex DNA (G4 DNA) in promoter regions of oncogenes and in telomeres has prompted the development of small molecules with high affinity and selectivity for these structures. Herein we report the synthesis of a new di-substituted bipyridine ligand and the corresponding complexes with Ni and VO . Both these new complexes have been characterized spectroscopically and by X-ray crystallography. Detailed DNA binding studies of these two complexes, together with three previously reported metal salphen complexes, are presented. Using FRET melting assays, the binding affinity and selectivity of the five metal complexes against six different G4 DNA structures as well as a duplex DNA have been determined. In addition, we present detailed ITC and UV/Vis studies to characterize the interaction of the complexes with human telomeric G4 DNA. Finally, we show via a polymerase stop assay that these complexes are able to stabilize a G4 DNA structure (from the c-Myc oncogene promoter) and halt the activity of Taq polymerase.

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Development of microfluidic platforms for the synthesis of metal complexes and evaluation of their DNA affinity using online FRET melting assays

Abstract: Two new microfluidic platforms are reported to synthesize metal complexes and assess their DNA affinity in continuous flow.

Cited by 18 publications

References 52 publications

G-Quadruplexes and Their Ligands: Biophysical Methods to Unravel G-Quadruplex/Ligand Interactions

G-Quadruplexes and Their Ligands: Biophysical Methods to Unravel G-Quadruplex/Ligand Interactions

An Octahedral Cobalt(III) Complex with Axial NH₃ Ligands that Templates and Selectively Stabilises G‐quadruplex DNA

Binding Studies of Metal–Salphen and Metal–Bipyridine Complexes towards G‐Quadruplex DNA

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Development of microfluidic platforms for the synthesis of metal complexes and evaluation of their DNA affinity using online FRET melting assays

Abstract: Two new microfluidic platforms are reported to synthesize metal complexes and assess their DNA affinity in continuous flow.

Cited by 18 publications

References 52 publications

G-Quadruplexes and Their Ligands: Biophysical Methods to Unravel G-Quadruplex/Ligand Interactions

G-Quadruplexes and Their Ligands: Biophysical Methods to Unravel G-Quadruplex/Ligand Interactions

An Octahedral Cobalt(III) Complex with Axial NH3 Ligands that Templates and Selectively Stabilises G‐quadruplex DNA

Binding Studies of Metal–Salphen and Metal–Bipyridine Complexes towards G‐Quadruplex DNA

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An Octahedral Cobalt(III) Complex with Axial NH₃ Ligands that Templates and Selectively Stabilises G‐quadruplex DNA