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

Dat Thinh Ha, Sthitodhi Ghosh, Chong H. Ahn, David Segal, Moon Soo Kim

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)4009-4016
Number of pages8
JournalAnalyst
Volume143
Issue number17
DOIs
StatePublished - Sep 7 2018

Fingerprint

Immobilized Proteins
Cycloparaffins
Zinc Fingers
Pathogens
Zinc
DNA sequences
Polymers
DNA
pathogen
polymer
zinc
Proteins
Lab-on-a-chip
protein
Copolymers
Olefins
Protein Array Analysis
Escherichia coli O157
Escherichia coli
Shiga Toxin

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Cite this

Pathogen-specific DNA sensing with engineered zinc finger proteins immobilized on a polymer chip. / Ha, Dat Thinh; Ghosh, Sthitodhi; Ahn, Chong H.; Segal, David; Kim, Moon Soo.

In: Analyst, Vol. 143, No. 17, 07.09.2018, p. 4009-4016.

Research output: Contribution to journalArticle

Ha, Dat Thinh ; Ghosh, Sthitodhi ; Ahn, Chong H. ; Segal, David ; Kim, Moon Soo. / Pathogen-specific DNA sensing with engineered zinc finger proteins immobilized on a polymer chip. In: Analyst. 2018 ; Vol. 143, No. 17. pp. 4009-4016.
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