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Pathogen-specific DNA sensing with engineered zinc finger proteins immobilized on polymer chip

Abstract

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 pathogen-specific DNA sequences present in E. coli O157:H7 on the lab-on-a chip. ZFPs custom-designed 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 with high specificity, providing a detection limit of 10 fmole of target DNA in colorimetric assay. Therefore, in this study, we demonstrated a great potential of ZFP arrays on the COC chip for further development of a simple and novel lab-on-a chip technology for detection of pathogens.

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Publication details

The article was received on 02 Mar 2018, accepted on 29 Jun 2018 and first published on 11 Jul 2018


Article type: Communication
DOI: 10.1039/C8AN00395E
Citation: Analyst, 2018, Accepted Manuscript
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    Pathogen-specific DNA sensing with engineered zinc finger proteins immobilized on polymer chip

    D. T. Ha , S. ghosh, C. H. Ahn, D. J. Segal and M. Kim, Analyst, 2018, Accepted Manuscript , DOI: 10.1039/C8AN00395E

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