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Aptamer Recognition of Multiplexed Small-Molecule-Functionalized Substrates.

Nako Nakatsuka, Huan H Cao, Stephanie Deshayes

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    This summary is machine-generated.

    Chemically synthesized DNA aptamers selectively recognize small-molecule targets like dopamine when immobilized on substrates. This method enables efficient aptamer selection and characterization for various applications.

    Keywords:
    chemical patterningfluorescence microscopymicrofluidicsmolecular recognitionneurotransmitteroligonucleotideself-assembled monolayers

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    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Chemical Biology

    Background:

    • Aptamers are nucleic acid or peptide molecules with high molecular recognition capabilities.
    • Traditional aptamer applications often focus on protein targets, with limited exploration of small-molecule interactions.
    • Developing methods for aptamer selection against small molecules is crucial for expanding their utility.

    Purpose of the Study:

    • To investigate the recognition of substrate-tethered small-molecule targets, specifically neurotransmitters, using DNA aptamers.
    • To establish a platform for the selective capture, sorting, and characterization of aptamers targeting small molecules.
    • To compare the binding affinity and selectivity of aptamers against various small molecules in situ.

    Main Methods:

    • Utilizing microfluidics to pattern small molecules (dopamine, l-tryptophan) onto substrate regions.
    • Employing fluorescently labeled DNA aptamers for detection and recognition of immobilized targets.
    • Synthesizing and using neurotransmitter-conjugated oligo(ethylene glycol) alkanethiols for creating multiplexed substrates.
    • Determining on-substrate dissociation constants and comparing them with solution-phase values.

    Main Results:

    • Demonstrated selective recognition of dopamine and l-tryptophan by their respective DNA aptamers when immobilized on substrates.
    • The on-substrate dissociation constant for the dopamine aptamer was comparable to its solution-phase value.
    • Successfully produced multiplexed substrates for aptamer capture and sorting using microfluidics and functionalized alkanethiols.
    • Enabled simultaneous determination of in situ binding constants for the dopamine aptamer against different analogues, revealing selectivity.

    Conclusions:

    • Strategically optimized small-molecule-functionalized substrates facilitate selective recognition by nucleic acid aptamers.
    • These substrates are valuable tools for side-by-side comparisons of aptamer binding and for the discovery of novel aptamers targeting small molecules.
    • The developed platform extends aptamer applications beyond protein targets to diverse small-molecule analytes.