Rolling Circular Amplification (RCA)-Assisted CRISPR/Cas9 Cleavage (RACE) for Highly Specific Detection of Multiple Extracellular Vesicle MicroRNAs
Multiplexed detection of extracellular vesicle (EV)-derived microRNAs (miRNAs) plays a critical role in facilitating disease diagnosis and prognosis evaluation. Herein, we developed a highly specific nucleic acid detection platform for simultaneous quantification of several EV-derived miRNAs in cons...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2020-01, Vol.92 (2), p.2176-2185 |
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| container_title | Analytical chemistry (Washington) |
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| creator | Wang, Ruixuan Zhao, Xianxian Chen, Xiaohui Qiu, Xiaopei Qing, Guangchao Zhang, Hong Zhang, Liangliang Hu, Xiaolin He, Zhuoqi Zhong, Daidi Wang, Ying Luo, Yang |
| description | Multiplexed detection of extracellular vesicle (EV)-derived microRNAs (miRNAs) plays a critical role in facilitating disease diagnosis and prognosis evaluation. Herein, we developed a highly specific nucleic acid detection platform for simultaneous quantification of several EV-derived miRNAs in constant temperature by integrating the advantages of a clustered regularly interspaced short palindromic repeats/CRISPR associated nucleases (CRISPR/Cas) system and rolling circular amplification (RCA) techniques. Particularly, the proposed approach demonstrated single-base resolution attributed to the dual-specific recognition from both padlock probe-mediated ligation and protospacer adjacent motif (PAM)-triggered cleavage. The high consistency between the proposed approach RCA-assisted CRISPR/Cas9 cleavage (RACE) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) in detecting EV-derived miRNAs’ abundance from both cultured cancer cells and clinical lung cancer patients validated its robustness, revealing its potentials in the screening, diagnosis, and prognosis of various diseases. In summary, RACE is a powerful tool for multiplexed, specific detection of nucleic acids in point-of-care diagnostics and field-deployable analysis. |
| doi_str_mv | 10.1021/acs.analchem.9b04814 |
| format | Article |
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Herein, we developed a highly specific nucleic acid detection platform for simultaneous quantification of several EV-derived miRNAs in constant temperature by integrating the advantages of a clustered regularly interspaced short palindromic repeats/CRISPR associated nucleases (CRISPR/Cas) system and rolling circular amplification (RCA) techniques. Particularly, the proposed approach demonstrated single-base resolution attributed to the dual-specific recognition from both padlock probe-mediated ligation and protospacer adjacent motif (PAM)-triggered cleavage. The high consistency between the proposed approach RCA-assisted CRISPR/Cas9 cleavage (RACE) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) in detecting EV-derived miRNAs’ abundance from both cultured cancer cells and clinical lung cancer patients validated its robustness, revealing its potentials in the screening, diagnosis, and prognosis of various diseases. In summary, RACE is a powerful tool for multiplexed, specific detection of nucleic acids in point-of-care diagnostics and field-deployable analysis.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.9b04814</identifier><identifier>PMID: 31875674</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>A549 Cells ; Amplification ; Chemistry ; Cleavage ; CRISPR ; CRISPR-Cas Systems - genetics ; Diagnosis ; Extracellular Vesicles - genetics ; Humans ; Lung cancer ; MicroRNAs ; MicroRNAs - blood ; MicroRNAs - genetics ; miRNA ; Multiplexing ; Nuclease ; Nucleic Acid Amplification Techniques ; Nucleic acids ; Polymerase chain reaction ; Prognosis ; Reverse Transcriptase Polymerase Chain Reaction ; Reverse transcription ; Temperature ; Tumor Cells, Cultured</subject><ispartof>Analytical chemistry (Washington), 2020-01, Vol.92 (2), p.2176-2185</ispartof><rights>Copyright American Chemical Society Jan 21, 2020</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a413t-4096abcbecd31f0b1127d56890c98e1a715f652e203591a2920e0842df6356e83</citedby><cites>FETCH-LOGICAL-a413t-4096abcbecd31f0b1127d56890c98e1a715f652e203591a2920e0842df6356e83</cites><orcidid>0000-0002-0166-9027</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/acs.analchem.9b04814$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.9b04814$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,777,781,2752,27057,27905,27906,56719,56769</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31875674$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Ruixuan</creatorcontrib><creatorcontrib>Zhao, Xianxian</creatorcontrib><creatorcontrib>Chen, Xiaohui</creatorcontrib><creatorcontrib>Qiu, Xiaopei</creatorcontrib><creatorcontrib>Qing, Guangchao</creatorcontrib><creatorcontrib>Zhang, Hong</creatorcontrib><creatorcontrib>Zhang, Liangliang</creatorcontrib><creatorcontrib>Hu, Xiaolin</creatorcontrib><creatorcontrib>He, Zhuoqi</creatorcontrib><creatorcontrib>Zhong, Daidi</creatorcontrib><creatorcontrib>Wang, Ying</creatorcontrib><creatorcontrib>Luo, Yang</creatorcontrib><title>Rolling Circular Amplification (RCA)-Assisted CRISPR/Cas9 Cleavage (RACE) for Highly Specific Detection of Multiple Extracellular Vesicle MicroRNAs</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Multiplexed detection of extracellular vesicle (EV)-derived microRNAs (miRNAs) plays a critical role in facilitating disease diagnosis and prognosis evaluation. Herein, we developed a highly specific nucleic acid detection platform for simultaneous quantification of several EV-derived miRNAs in constant temperature by integrating the advantages of a clustered regularly interspaced short palindromic repeats/CRISPR associated nucleases (CRISPR/Cas) system and rolling circular amplification (RCA) techniques. Particularly, the proposed approach demonstrated single-base resolution attributed to the dual-specific recognition from both padlock probe-mediated ligation and protospacer adjacent motif (PAM)-triggered cleavage. The high consistency between the proposed approach RCA-assisted CRISPR/Cas9 cleavage (RACE) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) in detecting EV-derived miRNAs’ abundance from both cultured cancer cells and clinical lung cancer patients validated its robustness, revealing its potentials in the screening, diagnosis, and prognosis of various diseases. 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Zhao, Xianxian ; Chen, Xiaohui ; Qiu, Xiaopei ; Qing, Guangchao ; Zhang, Hong ; Zhang, Liangliang ; Hu, Xiaolin ; He, Zhuoqi ; Zhong, Daidi ; Wang, Ying ; Luo, Yang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a413t-4096abcbecd31f0b1127d56890c98e1a715f652e203591a2920e0842df6356e83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>A549 Cells</topic><topic>Amplification</topic><topic>Chemistry</topic><topic>Cleavage</topic><topic>CRISPR</topic><topic>CRISPR-Cas Systems - genetics</topic><topic>Diagnosis</topic><topic>Extracellular Vesicles - genetics</topic><topic>Humans</topic><topic>Lung cancer</topic><topic>MicroRNAs</topic><topic>MicroRNAs - blood</topic><topic>MicroRNAs - genetics</topic><topic>miRNA</topic><topic>Multiplexing</topic><topic>Nuclease</topic><topic>Nucleic Acid Amplification Techniques</topic><topic>Nucleic acids</topic><topic>Polymerase chain reaction</topic><topic>Prognosis</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Reverse transcription</topic><topic>Temperature</topic><topic>Tumor Cells, Cultured</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Ruixuan</creatorcontrib><creatorcontrib>Zhao, Xianxian</creatorcontrib><creatorcontrib>Chen, Xiaohui</creatorcontrib><creatorcontrib>Qiu, Xiaopei</creatorcontrib><creatorcontrib>Qing, Guangchao</creatorcontrib><creatorcontrib>Zhang, Hong</creatorcontrib><creatorcontrib>Zhang, Liangliang</creatorcontrib><creatorcontrib>Hu, Xiaolin</creatorcontrib><creatorcontrib>He, Zhuoqi</creatorcontrib><creatorcontrib>Zhong, Daidi</creatorcontrib><creatorcontrib>Wang, Ying</creatorcontrib><creatorcontrib>Luo, Yang</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Ruixuan</au><au>Zhao, Xianxian</au><au>Chen, Xiaohui</au><au>Qiu, Xiaopei</au><au>Qing, Guangchao</au><au>Zhang, Hong</au><au>Zhang, Liangliang</au><au>Hu, Xiaolin</au><au>He, Zhuoqi</au><au>Zhong, Daidi</au><au>Wang, Ying</au><au>Luo, Yang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rolling Circular Amplification (RCA)-Assisted CRISPR/Cas9 Cleavage (RACE) for Highly Specific Detection of Multiple Extracellular Vesicle MicroRNAs</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2020-01-21</date><risdate>2020</risdate><volume>92</volume><issue>2</issue><spage>2176</spage><epage>2185</epage><pages>2176-2185</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>Multiplexed detection of extracellular vesicle (EV)-derived microRNAs (miRNAs) plays a critical role in facilitating disease diagnosis and prognosis evaluation. Herein, we developed a highly specific nucleic acid detection platform for simultaneous quantification of several EV-derived miRNAs in constant temperature by integrating the advantages of a clustered regularly interspaced short palindromic repeats/CRISPR associated nucleases (CRISPR/Cas) system and rolling circular amplification (RCA) techniques. Particularly, the proposed approach demonstrated single-base resolution attributed to the dual-specific recognition from both padlock probe-mediated ligation and protospacer adjacent motif (PAM)-triggered cleavage. The high consistency between the proposed approach RCA-assisted CRISPR/Cas9 cleavage (RACE) and reverse transcription quantitative polymerase chain reaction (RT-qPCR) in detecting EV-derived miRNAs’ abundance from both cultured cancer cells and clinical lung cancer patients validated its robustness, revealing its potentials in the screening, diagnosis, and prognosis of various diseases. In summary, RACE is a powerful tool for multiplexed, specific detection of nucleic acids in point-of-care diagnostics and field-deployable analysis.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>31875674</pmid><doi>10.1021/acs.analchem.9b04814</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0002-0166-9027</orcidid></addata></record> |
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| subjects | A549 Cells Amplification Chemistry Cleavage CRISPR CRISPR-Cas Systems - genetics Diagnosis Extracellular Vesicles - genetics Humans Lung cancer MicroRNAs MicroRNAs - blood MicroRNAs - genetics miRNA Multiplexing Nuclease Nucleic Acid Amplification Techniques Nucleic acids Polymerase chain reaction Prognosis Reverse Transcriptase Polymerase Chain Reaction Reverse transcription Temperature Tumor Cells, Cultured |
| title | Rolling Circular Amplification (RCA)-Assisted CRISPR/Cas9 Cleavage (RACE) for Highly Specific Detection of Multiple Extracellular Vesicle MicroRNAs |
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