Pathogen-specific DNA sensing with engineered zinc finger proteins immobilized on a polymer chip. Issue 17 (25th July 2018)

• Record Type: Journal Article
• Authors: Ha, Dat Thinh; Ghosh, Sthitodhi; Ahn, Chong H.; Segal, David J.; Kim, Moon-Soo
• Is Part Of: Analyst. Volume 143:Issue 17(2018)
• Page Start: 4009
• Page End: 4016
• Publication Date: 2018-07-25
• Subjects: Chemistry, Analytic -- Periodicals; 543
• DOI: 10.1039/c8an00395e
• Languages: English
• ISSNs: 0003-2654

Abstract

Abstract : An array of engineered zinc finger proteins (ZFPs) along with the detection ZFP probes for sensing pathogen-specific DNA on a cyclic olefin copolymer 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 the recognition of specific DNA sequences. As a proof of concept, a sequence-enabled reassembly of a 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 a lab-on-a-chip. ZFPs custom-designed to detect Shiga toxin in E. coli O157:H7 were immobilized on a cyclic olefin copolymer (COC) chip, which can function as a non-PCR based molecular diagnostic device. Pathogen-specific double-stranded DNA was directly detected by using engineered ZFPs immobilized on the COC chip with high specificity, providing a detection limit of 10 fmol of target DNA in a colorimetric assay. Therefore, in this study, we demonstrated the great potential of ZFP arrays on the COC chip for further development of a simple and novel lab-on-a-chip technology for the detection of pathogens.