Magnetic bead-based electrochemical assay for determination of DNA methyltransferase activity

Dudová, Zdenka; Bartošík, Martin; Fojta, Miroslav

doi:10.1016/j.electacta.2017.02.104

Cited by 10 publications

(3 citation statements)

References 29 publications

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“…Next, we aimed to generate self-DNA with a different content of methylated CpG motifs. To this end, we pre-treated bean self-DNA with the DNA methyltransferase M.SssI, which methylates all cytosine residues in the 5'-CG-3' motif 39 , and subsequently subjected fractions of natural and M.SssI-methylated DNA to digestion with each of two restriction enzymes, MspI and HpaII, which recognize 5′-CCGG-3′ motifs and cleave between the two cytosines [40][41][42] (see Fig. S5 for a graphical illustration).…”

Section: Resultsmentioning

confidence: 99%

“…S5 for a graphical illustration). Both enzymes can cleave unmethylated 5′-CCGG-3′ motifs and neither of them can cleave when the external cytosine residue is methylated (5′-m CCGG-3′), but MspI can cleave when the internal cytosine is methylated (5′-C m CGG-3′), whereas HpaII cannot [40][41][42] . Correspondingly, pre-treatment with M.SssI protected genomic DNA of Aspergillus fumigatus from digestion by HpaII, but not MspI, and pre-treatment with each of these enzymes reduced the effects of microbial DNA on cytokine production by mouse bone marrow-derived dendritic cells 43 : both responses considered as canonical TLR9-dependent mammalian immune responses to non-self-DNA 11 .…”

Section: Resultsmentioning

confidence: 99%

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The CpG-dependent plant immune response to self-DNA triggers defence hormone signalling and improves fitness

Durán-Flores

Heil

2023

Preprint

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The accumulation of DNA in the cytoplasm or extracellular space is a signal of danger. Plants respond to this signal with a self/non-self-specific activation of early immune signalling events. Here, we asked whether this specificity translates to fitness-relevant resistance to natural enemies. We treated common bean (Phaseolus vulgaris) plants with self-DNA and with non-self-DNA from other plant species. Self-DNA treatment induced jasmonic acid and decreased feeding by a chewing herbivore (Spodoptera frugiperda), while self- and non-self-DNA induced salicylic acid and reduced the population densities of two fungal pathogens (Botrytis cinerea and Sclerotinia sclerotiorum) and of four bacterial pathogens (Enterobacter sp. strain FCB1, Pseudomonas syringae pv. phaseoli and pv. syringae, and Xanthomonas axonopodis pv. phaseoli). Strikingly, a single self-DNA-treatment increased seed production under field conditions in two seasons ca. 1.5-fold and 3.2-fold, while non-self-DNA had lower or no detectable effects. Stronger responses to self- than non-self-DNA seemingly contradict immunological theory. In mammalian immune cells, toll-like receptor 9 (TLR9) is activated by microbial DNA but also by mitochondrial (mt)DNA that is rich in unmethylated CpG motifs. Using bean suspension cells, we observed stronger immunogenic effects of genomic DNA than chloroplast DNA or mtDNA. Moreover, in vitro methylation or cleavage of CpG motifs reduced – without eliminating – the H2O2-inducing properties of DNA. We conclude that the plant immune response to DNA comprises a self/non-self-specific induction of major defence hormones that can be adaptive under natural enemy pressure and that unmethylated CpG motifs contribute to the immunogenic effects of DNA in plants.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

The CpG-dependent plant immune response to self-DNA triggers defence hormone signalling and improves fitness

Durán-Flores

Heil

2023

Preprint

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“…Magnetic nanoparticles (MNPs) are appropriate platforms for such bioassays due to their easy functionalization through the attachment of organic or biological molecules to the MNP surface, which increases the sensitivity and bonding strength of the target to the solid support [15]. Moreover, the potential of their magnetic properties to perform their separation from solvents and other chemicals, eliminating tedious and costly separation processes [16], making these nanomaterials a good choice for the development of analytical protocols that are faster, simpler and more precise that existing methodologies [17].…”

Section: Introductionmentioning

confidence: 99%

Fe3O4@Au nanoparticles-based magnetoplatform for the HMGA maize endogenous gene electrochemical genosensing

Sousa

Ramos‐Jesus²,

Silva

et al. 2020

Talanta

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This work addresses a technological advance applied to the construction of a magnetogenoassay with electrochemical transduction for the maize taxon-specific (HMGA gene) detection using gold-coated magnetic nanoparticles as nanosized platform.Superparamagnetic core-shell Fe3O4@Au nanoparticles (10.4 ± 1.7 nm) were used to assemble the genoassay through the covalent immobilization of HMGA DNA probes onto carboxylated self-assembled monolayers at the nanoparticles surface.A hybridization reaction using sandwich format was selected to prevent inefficient hybridization connected with stable secondary DNA structures using also fluorescein isothiocyanate as DNA signaling tag. The labelling of the hybridization reaction with enzymes allowed the chronoamperometric measurement of the peroxidase activity linked to the nanoplatform located on gold surface.Using this electrochemical magnetogenoassay a linear concentration range from 0.5 to 5 nM and a LOD of 90 pM with a RSD<1.2% was calculated. Certified maize was evaluated without further purification after PCR amplification. This work highlights the efficacy of the electrochemical magnetogenoassay for the HMGA detection, showing its potential as alternative procedure for the verification of the compliance of the legislation.

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Amplification strategy for sensitive detection of methyltransferase activity based on surface plasma resonance techniques

Zhang

Liang

et al. 2018

Analytica Chimica Acta

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Magnetic bead-based electrochemical assay for determination of DNA methyltransferase activity

Cited by 10 publications

References 29 publications

The CpG-dependent plant immune response to self-DNA triggers defence hormone signalling and improves fitness

The CpG-dependent plant immune response to self-DNA triggers defence hormone signalling and improves fitness

Fe3O4@Au nanoparticles-based magnetoplatform for the HMGA maize endogenous gene electrochemical genosensing

Amplification strategy for sensitive detection of methyltransferase activity based on surface plasma resonance techniques

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