Specific DNA recognition is of great importance for clinical and biomedical applications as well as public health. Emerging applications include detecting cancer biomarkers, pathogens, and antibiotic resistant genes (ARGs). Zinc finger proteins (ZFPs) are the most abundant DNA binding domain in eukaryotic genomes. ZFPs provide a versatile scaffold to design for direct detection of double-stranded (ds) DNA. In this study, a sandwich assay for detecting specific ARGs has been demonstrated by employing a pair of ZFPs which consists of capture and detection probes. The capture probe ZFP was conjugated on the silane polymer-coated surface through APDMES (3-aminopropyldimethylethoxysilane) cross-linking, creating an anticipated self-assembled monolayer with the capture probes conjugated homogeneously. The APDMES-functionalized surface was utilized as a platform for detection of specific dsDNA. The organic dye-labeled ZFP was used as the detection probe in this assay, generating the fluorescence signals from the bound complex of capture probes, dsDNA, and detection probes upon ZFPs binding to target DNA.  We have demonstrated for the first time that the APDMES monolayer surface can be used to detect dsDNA along with conjugated DNA-binding proteins.

Engineered zinc finger proteins immobilized on the silane polymer surface for diagnostic probes to  detect antibiotic resistance genes

2019 Spring National Meeting

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