A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications
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 rea...
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| Veröffentlicht in: | Nucleic acids research 2011-03, Vol.39 (5), p.e29-e29 |
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| container_end_page | e29 |
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| container_issue | 5 |
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| container_title | Nucleic acids research |
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| creator | Kim, Moon-Soo Stybayeva, Gulnaz Lee, Ji Youn Revzin, Alexander Segal, David J |
| description | 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 50 fmol of target DNA. The method could distinguish target DNA from non-target DNA within 5 min. 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. |
| doi_str_mv | 10.1093/nar/gkq1214 |
| format | Article |
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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 50 fmol of target DNA. The method could distinguish target DNA from non-target DNA within 5 min. 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.</description><subject>beta-Lactamases - genetics</subject><subject>DNA, Bacterial - chemistry</subject><subject>DNA-Binding Proteins - analysis</subject><subject>DNA-Binding Proteins - genetics</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Escherichia coli - genetics</subject><subject>Methods Online</subject><subject>Molecular Diagnostic Techniques</subject><subject>Protein Array Analysis - methods</subject><subject>Protein Engineering</subject><subject>Sequence Analysis, DNA</subject><subject>Zinc Fingers</subject><issn>0305-1048</issn><issn>1362-4962</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkTFvFDEQhS0EIkegokfuKNASj-312Q3SKYEkUkQaqC2fPb447Nkbey9S-PXZJEcEFdUU87038_QIeQ_sMzAjjrKrR5tfN8BBviALEIp30ij-kiyYYH0HTOoD8qa1a8ZAQi9fkwMOIKRhZkGuV_R3yp7GlDdY6VjLhClTV6u7o7FUOl0hvU1t5wYacEI_pZJpibSN6FNMnp58X9GGNzvMHtujJCS3yaVN89KN45C8exC1t-RVdEPDd_t5SH5--_rj-Ky7uDw9P15ddF4u9dQFE6TRa8N70QummOBSrZkJSvVzXMeXTqtodAwycCEQes5MjFpKFkB57MUh-fLkO-7WWwwe81TdYMeatq7e2eKS_XeT05XdlFs7XwOmzGzwcW9Qy5yrTXabmsdhcBnLrlltDCgAs_w_2WuujVJiJj89kb6W1irG53-A2Yca7Vyj3dc40x_-jvDM_ulN3AM7jpqX</recordid><startdate>20110301</startdate><enddate>20110301</enddate><creator>Kim, Moon-Soo</creator><creator>Stybayeva, Gulnaz</creator><creator>Lee, Ji Youn</creator><creator>Revzin, Alexander</creator><creator>Segal, David J</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7QL</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>20110301</creationdate><title>A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications</title><author>Kim, Moon-Soo ; Stybayeva, Gulnaz ; Lee, Ji Youn ; Revzin, Alexander ; Segal, David J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c478t-d9d498b9253530603246b09d665109a27a86f98fd4d233e15209ff8440d16ce53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>beta-Lactamases - genetics</topic><topic>DNA, Bacterial - chemistry</topic><topic>DNA-Binding Proteins - analysis</topic><topic>DNA-Binding Proteins - genetics</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Escherichia coli - genetics</topic><topic>Methods Online</topic><topic>Molecular Diagnostic Techniques</topic><topic>Protein Array Analysis - methods</topic><topic>Protein Engineering</topic><topic>Sequence Analysis, DNA</topic><topic>Zinc Fingers</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kim, Moon-Soo</creatorcontrib><creatorcontrib>Stybayeva, Gulnaz</creatorcontrib><creatorcontrib>Lee, Ji Youn</creatorcontrib><creatorcontrib>Revzin, Alexander</creatorcontrib><creatorcontrib>Segal, David J</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Nucleic acids research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kim, Moon-Soo</au><au>Stybayeva, Gulnaz</au><au>Lee, Ji Youn</au><au>Revzin, Alexander</au><au>Segal, David J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications</atitle><jtitle>Nucleic acids research</jtitle><addtitle>Nucleic Acids Res</addtitle><date>2011-03-01</date><risdate>2011</risdate><volume>39</volume><issue>5</issue><spage>e29</spage><epage>e29</epage><pages>e29-e29</pages><issn>0305-1048</issn><eissn>1362-4962</eissn><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 50 fmol of target DNA. The method could distinguish target DNA from non-target DNA within 5 min. The ZFP arrays provided sufficient sensitivity and high specificity to recognize specific DNA sequences. 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| source | MEDLINE; DOAJ Directory of Open Access Journals; Oxford Journals Open Access Collection; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | beta-Lactamases - genetics DNA, Bacterial - chemistry DNA-Binding Proteins - analysis DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism Escherichia coli - genetics Methods Online Molecular Diagnostic Techniques Protein Array Analysis - methods Protein Engineering Sequence Analysis, DNA Zinc Fingers |
| title | A zinc finger protein array for the visual detection of specific DNA sequences for diagnostic applications |
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