DNA‐Assemblierung mittels Affinitätsbindung für die ultraempfindliche Proteindetektion

Zhang, Hongquan; Feng, Lu; Dever, Brittany; Wang, Chuan; Li, Xing‐Fang; Le, X. Chris

doi:10.1002/ange.201210022

Cited by 17 publications

(1 citation statement)

References 58 publications

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“…Similar concept of binding-induced DNA assembly was recently reported by Le's group where the co-binding of two reporter molecules on a single protein molecule increased the local probe concentration and drastically enhanced the kinetic rate of strand exchange ( 14 , 15 ). While their proposed format works well for homogeneous assay ( 16 , 17 ), the fluorescence signal developed is freely moving in the bulk solution, and render it difficult to extend the design to pinpoint and visualize the site of interaction. Therefore, we aim to engineer a self-contained DNA circuit on the binding site itself where the localized readout signal is developed and amplified.…”

Section: Introductionmentioning

confidence: 99%

Engineering self-contained DNA circuit for proximity recognition and localized signal amplification of target biomolecules

Ang

Yung

2014

Nucleic Acids Res

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Biomolecular interactions have important cellular implications, however, a simple method for the sensing of such proximal events is lacking in the current molecular toolbox. We designed a dynamic DNA circuit capable of recognizing targets in close proximity to initiate a pre-programmed signal transduction process resulting in localized signal amplification. The entire circuit was engineered to be self-contained, i.e. it can self-assemble onto individual target molecules autonomously and form localized signal with minimal cross-talk. α-thrombin was used as a model protein to evaluate the performance of the individual modules and the overall circuit for proximity interaction under physiologically relevant buffer condition. The circuit achieved good selectivity in presence of non-specific protein and interfering serum matrix and successfully detected for physiologically relevant α-thrombin concentration (50 nM–5 μM) in a single mixing step without any further washing. The formation of localized signal at the interaction site can be enhanced kinetically through the control of temperature and probe concentration. This work provides a basic general framework from which other circuit modules can be adapted for the sensing of other biomolecular or cellular interaction of interest.

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Section: Introductionmentioning

confidence: 99%

Engineering self-contained DNA circuit for proximity recognition and localized signal amplification of target biomolecules

Ang

Yung

2014

Nucleic Acids Res

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Binding‐Induced DNA Nanomachines Triggered by Proteins and Nucleic Acids

et al. 2015

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We introduce the concept and operation of ab inding-induced DNAn anomachine that can be activated by proteins and nucleic acids.T his new type of nanomachine harnesses specific target binding to trigger assembly of separate DNAc omponents that are otherwise unable to spontaneously assemble.T hree-dimensional DNAt racks of high density are constructed on gold nanoparticles functionalizedw ith hundreds of single-stranded oligonucleotides and tens of an affinity ligand. AD NA swing arm, free in solution, is linked to asecond affinity ligand. Binding of atarget molecule to the two ligands brings the swing arm to AuNP and initiates autonomous,stepwise movement of the swing arm around the AuNP surface.T he movemento ft he swing arm, powered by enzymatic cleavage of conjugated oligonucleotides,c leaves hundreds of oligonucleotides in response to as ingle binding event. We demonstrate three nanomachines that are specifically activated by streptavidin, platelet-derived growth factor,a nd the Smallpox gene.S ubstituting the ligands enables the nanomachine to respond to other molecules.The new nanomachines have several unique and advantageous features over DNA nanomachines that rely on DNAs elf-assembly.

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Target‐Induced and Equipment‐Free DNA Amplification with a Simple Paper Device

Hui

Zhang

et al. 2016

Angewandte Chemie

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We report on a paper device capable of carrying out target‐induced rolling circle amplification (RCA) to produce massive DNA amplicons that can be easily visualized. Interestingly, we observed that RCA was more proficient on paper than in solution, which we attribute to a significantly higher localized concentration of immobilized DNA. Furthermore, we have successfully engineered a fully functional paper device for sensitive DNA or microRNA detection via printing of all RCA‐enabling molecules within a polymeric sugar film formed from pullulan, which was integrated with the paper device. This encapsulation not only stabilizes the entrapped reagents at room temperature but also enables colorimetric bioassays with minimal steps.

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DNA‐Assemblierung mittels Affinitätsbindung für die ultraempfindliche Proteindetektion

Cited by 17 publications

References 58 publications

Engineering self-contained DNA circuit for proximity recognition and localized signal amplification of target biomolecules

Engineering self-contained DNA circuit for proximity recognition and localized signal amplification of target biomolecules

Binding‐Induced DNA Nanomachines Triggered by Proteins and Nucleic Acids

Target‐Induced and Equipment‐Free DNA Amplification with a Simple Paper Device

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