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An Engineered Split-TET2 Enzyme for Chemical-inducible DNA Hydroxymethylation and Epigenetic Remodeling
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    This study applies CMOS design to ion-sensitive field-effect transistors (ISFETs) for DNA methylation and miRNA detection. Novel circuits improve accuracy and speed for point-of-care diagnostics and portable real-time monitoring.

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    Area of Science:

    • Biomedical Engineering
    • Electronics Engineering
    • Molecular Biology

    Background:

    • Ion-sensitive field-effect transistors (ISFETs) are crucial for biosensing.
    • CMOS design offers a scalable platform for integrated biosensor development.
    • Accurate detection of DNA methylation and microRNAs (miRNAs) is vital for diagnostics.

    Purpose of the Study:

    • To explore the application of CMOS design methods for ISFET-based DNA methylation and miRNA detection.
    • To develop novel front-end analog and back-end digital circuits for enhanced biosensing capabilities.
    • To improve the accuracy, speed, and portability of molecular diagnostic platforms.

    Main Methods:

    • Design of ISFET weak inversion current mirror topology for differential sensing in Methylation-specific PCR (MSP).
    • Implementation of a switched current integrator to enhance accuracy in ion-semiconductor sequencing.
    • Development of two threshold cycle evaluation systems (Derivative and 3-point exponential) for real-time miRNA quantification.
    • Fabrication of all designs in unmodified CMOS technology.

    Main Results:

    • Demonstrated ISFET topology for sensitive and stable differential sensing.
    • Showcased potential for improved homopolymer detection accuracy using switched current integrators.
    • Validated real-time monitoring systems for DNA amplification with reduced detection times.
    • Assessed performance based on functionality and practical pH-resolution requirements.

    Conclusions:

    • CMOS design methods provide a robust framework for developing advanced ISFET-based biosensors.
    • The developed circuits offer improved performance for DNA methylation and miRNA detection.
    • This work paves the way for highly integrated, portable, and efficient point-of-care diagnostic devices.