A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications

Moon Soo Kim; Gulnaz Stybayeva; Ji Youn Lee; Alexander Revzin; David J. Segal

doi:10.1093/nar/gkq1214

A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications

Moon Soo Kim, Gulnaz Stybayeva, Ji Youn Lee, Alexander Revzin, David J. Segal

Physiology & Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

The visual detection of specific double-stranded DNA sequences possesses great potential for the development of diagnostics. Zinc finger domains provide a powerful scaffold for creating custom DNA-binding proteins that recognize specific DNA sequences. We previously demonstrated sequence-enabled reassembly of TEM-1 β-lactamase (SEER-LAC), a system consisting of two inactive fragments of β-lactamase each linked to engineered zinc finger proteins (ZFPs). Here the SEER-LAC system was applied to develop ZFP arrays that function as simple devices to identify bacterial double-stranded DNA sequences. The ZFP arrays provided a quantitative assay with a detection limit of 50fmol of target DNA. The method could distinguish target DNA from non-target DNA within 5min. The ZFP arrays provided sufficient sensitivity and high specificity to recognize specific DNA sequences. These results suggest that ZFP arrays have the potential to be developed into a simple and rapid point-of-care (POC) diagnostic for the multiplexed detection of pathogens.

Original language	English (US)
Pages (from-to)	e29
Journal	Nucleic acids research
Volume	39
Issue number	5
DOIs	https://doi.org/10.1093/nar/gkq1214
State	Published - Mar 2011

ASJC Scopus subject areas

Genetics

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abstract = "The visual detection of specific double-stranded DNA sequences possesses great potential for the development of diagnostics. Zinc finger domains provide a powerful scaffold for creating custom DNA-binding proteins that recognize specific DNA sequences. We previously demonstrated sequence-enabled reassembly of TEM-1 β-lactamase (SEER-LAC), a system consisting of two inactive fragments of β-lactamase each linked to engineered zinc finger proteins (ZFPs). Here the SEER-LAC system was applied to develop ZFP arrays that function as simple devices to identify bacterial double-stranded DNA sequences. The ZFP arrays provided a quantitative assay with a detection limit of 50fmol of target DNA. The method could distinguish target DNA from non-target DNA within 5min. The ZFP arrays provided sufficient sensitivity and high specificity to recognize specific DNA sequences. These results suggest that ZFP arrays have the potential to be developed into a simple and rapid point-of-care (POC) diagnostic for the multiplexed detection of pathogens.",

author = "Kim, {Moon Soo} and Gulnaz Stybayeva and Lee, {Ji Youn} and Alexander Revzin and Segal, {David J.}",

note = "Funding Information: National Institutes of Health (GM077403 to D.J.S.); National Institutes of Health, University of California, Davis-Lawrence Livermore National Laboratories Point-of-Care Technologies Center (pilot grant EB007959). Funding for open access charge: National Institutes of Health (GM077403).",

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AU - Revzin, Alexander

AU - Segal, David J.

N1 - Funding Information: National Institutes of Health (GM077403 to D.J.S.); National Institutes of Health, University of California, Davis-Lawrence Livermore National Laboratories Point-of-Care Technologies Center (pilot grant EB007959). Funding for open access charge: National Institutes of Health (GM077403).

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A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications

Abstract

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