Developing a New Sensing Technology for Double-Stranded DNA Detection Utilizing Engineered Zinc Finger Proteins and Nanomaterials (original) (raw)
A specific double-stranded DNA sensing system is of great interest for diagnostic and other biomedical applications. Zinc finger domains, which recognize double-stranded DNA, can be engineered to form custom DNA-binding proteins for recognition of specific DNA sequences. As a proof of concept, a sequence-enabled reassembly of TEM-1 β- lactamase system (SEER-LAC) was previously demonstrated to develop zinc finger protein (ZFP) arrays for the detection of a double-stranded bacterial DNA sequence. Here, we implemented the SEER-LAC system to demonstrate the direct detection of pathogenspecific DNA sequences present in E. coli O157:H7 on the lab-on-a chip. ZFPs customdesigned to detect shiga toxin in E. coli O157:H7 were immobilized on the cyclic olefin copolymer (COC) chip, which can function as a non-PCR based molecular diagnostic. Pathogen-specific double-stranded DNA was directly detected by engineered ZFPs immobilized on the COC chip, providing a detection limit of 10 fmole of targe...
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