CRISPR-Cas-Driven Single Micromotor (Cas-DSM) Enables Direct Detection of Nucleic Acid Biomarkers at the Single-Molecule Level
The target-dependent endonuclease activity (also known as the trans-cleavage activity) of CRISPR-Cas systems has stimulated great interest in the development of nascent sensing strategies for nucleic acid diagnostics. Despite many attempts, the majority of the sensitive CRISPR-Cas diagnostics strate...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2023-04, Vol.95 (13), p.5729-5737 |
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| creator | Chen, Desheng Liang, Yuanwen Wang, Honghong Wang, Hui Su, Fengxia Zhang, Pengbo Wang, Shuhui Liu, Weiliang Li, Zhengping |
| description | The target-dependent endonuclease activity (also known as the trans-cleavage activity) of CRISPR-Cas systems has stimulated great interest in the development of nascent sensing strategies for nucleic acid diagnostics. Despite many attempts, the majority of the sensitive CRISPR-Cas diagnostics strategies mainly rely on nucleic acid preamplification, which generally needs complex probes/primers designs, multiple experimental steps, and a longer testing time, as well as introducing the risk of false-positive results. In this work, we propose the CRISPR-Cas-Driven Single Micromotor (Cas-DSM), which can directly detect the nucleic acid targets at a single-molecule level with high specificity. We have demonstrated that the Cas-DSM is a reliable and practical method for the quantitative detection of DNA/RNA in various complex clinical samples as well as in individual cells without any preamplification processes. Due to the excellent features of the CRISPR/Cas system, including constant temperature, simple design, high specificity, and flexible programmability, the Cas-DSM could serve as a simple and universal platform for nucleic acid detection. More importantly, this work will provide a breakthrough for the development of next-generation amplification-free CRISPR/Cas sensing toolboxes. |
| doi_str_mv | 10.1021/acs.analchem.2c05767 |
| format | Article |
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Despite many attempts, the majority of the sensitive CRISPR-Cas diagnostics strategies mainly rely on nucleic acid preamplification, which generally needs complex probes/primers designs, multiple experimental steps, and a longer testing time, as well as introducing the risk of false-positive results. In this work, we propose the CRISPR-Cas-Driven Single Micromotor (Cas-DSM), which can directly detect the nucleic acid targets at a single-molecule level with high specificity. We have demonstrated that the Cas-DSM is a reliable and practical method for the quantitative detection of DNA/RNA in various complex clinical samples as well as in individual cells without any preamplification processes. Due to the excellent features of the CRISPR/Cas system, including constant temperature, simple design, high specificity, and flexible programmability, the Cas-DSM could serve as a simple and universal platform for nucleic acid detection. More importantly, this work will provide a breakthrough for the development of next-generation amplification-free CRISPR/Cas sensing toolboxes.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/acs.analchem.2c05767</identifier><identifier>PMID: 36944919</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Acids ; Biomarkers ; Biosensing Techniques ; Chemistry ; CRISPR ; CRISPR-Cas Systems - genetics ; DNA Primers ; DNA probes ; Endonuclease ; Micromotors ; Nucleic Acids ; RNA ; Target detection</subject><ispartof>Analytical chemistry (Washington), 2023-04, Vol.95 (13), p.5729-5737</ispartof><rights>2023 American Chemical Society</rights><rights>Copyright American Chemical Society Apr 4, 2023</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a376t-dd9cde623ad78b37f2a827b5cfcc711481b2bb2ce5244061eeaab77151fd58363</citedby><cites>FETCH-LOGICAL-a376t-dd9cde623ad78b37f2a827b5cfcc711481b2bb2ce5244061eeaab77151fd58363</cites><orcidid>0000-0002-8535-5358 ; 0000-0002-5116-1977 ; 0000-0002-4281-0541 ; 0000-0002-7573-8822</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.2c05767$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/acs.analchem.2c05767$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,776,780,2751,27055,27903,27904,56717,56767</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/36944919$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chen, Desheng</creatorcontrib><creatorcontrib>Liang, Yuanwen</creatorcontrib><creatorcontrib>Wang, Honghong</creatorcontrib><creatorcontrib>Wang, Hui</creatorcontrib><creatorcontrib>Su, Fengxia</creatorcontrib><creatorcontrib>Zhang, Pengbo</creatorcontrib><creatorcontrib>Wang, Shuhui</creatorcontrib><creatorcontrib>Liu, Weiliang</creatorcontrib><creatorcontrib>Li, Zhengping</creatorcontrib><title>CRISPR-Cas-Driven Single Micromotor (Cas-DSM) Enables Direct Detection of Nucleic Acid Biomarkers at the Single-Molecule Level</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>The target-dependent endonuclease activity (also known as the trans-cleavage activity) of CRISPR-Cas systems has stimulated great interest in the development of nascent sensing strategies for nucleic acid diagnostics. Despite many attempts, the majority of the sensitive CRISPR-Cas diagnostics strategies mainly rely on nucleic acid preamplification, which generally needs complex probes/primers designs, multiple experimental steps, and a longer testing time, as well as introducing the risk of false-positive results. In this work, we propose the CRISPR-Cas-Driven Single Micromotor (Cas-DSM), which can directly detect the nucleic acid targets at a single-molecule level with high specificity. We have demonstrated that the Cas-DSM is a reliable and practical method for the quantitative detection of DNA/RNA in various complex clinical samples as well as in individual cells without any preamplification processes. Due to the excellent features of the CRISPR/Cas system, including constant temperature, simple design, high specificity, and flexible programmability, the Cas-DSM could serve as a simple and universal platform for nucleic acid detection. More importantly, this work will provide a breakthrough for the development of next-generation amplification-free CRISPR/Cas sensing toolboxes.</description><subject>Acids</subject><subject>Biomarkers</subject><subject>Biosensing Techniques</subject><subject>Chemistry</subject><subject>CRISPR</subject><subject>CRISPR-Cas Systems - genetics</subject><subject>DNA Primers</subject><subject>DNA probes</subject><subject>Endonuclease</subject><subject>Micromotors</subject><subject>Nucleic Acids</subject><subject>RNA</subject><subject>Target detection</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1P3DAQhq2qqCy0_6BClnqBQxaPk9jJEXb5knbbim3PkeNMwNSJwU6QuPDb8bILhx44zWGe97U8DyHfgU2BcThWOkxVr6y-xW7KNculkJ_IBHLOElEU_DOZMMbShEvGdsleCHeMATAQX8huKsosK6GckOfZ9dXq93UyUyGZe_OIPV2Z_sYiXRrtXecG5-nh63a1PKJnvaotBjo3HvVA5zjEYVxPXUt_jtqi0fREm4aeGtcp_w99oGqgwy1ua5Ols6jH2L_AR7RfyU6rbMBv27lP_p6f_ZldJotfF1ezk0WiUimGpGlK3aDgqWpkUaey5argss51q7UEyAqoeV1zjTnPMiYAUalaSsihbfIiFek-Odz03nv3MGIYqs4EjdaqHt0YKi6LUgLPcojoj__QOzf6eOg1VZacZ1Kkkco2VDxSCB7b6t6b-OOnCli19lNFP9Wbn2rrJ8YOtuVj3WHzHnoTEgG2Adbx94c_7HwBmsWePQ</recordid><startdate>20230404</startdate><enddate>20230404</enddate><creator>Chen, Desheng</creator><creator>Liang, Yuanwen</creator><creator>Wang, Honghong</creator><creator>Wang, Hui</creator><creator>Su, Fengxia</creator><creator>Zhang, Pengbo</creator><creator>Wang, Shuhui</creator><creator>Liu, Weiliang</creator><creator>Li, Zhengping</creator><general>American Chemical Society</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-8535-5358</orcidid><orcidid>https://orcid.org/0000-0002-5116-1977</orcidid><orcidid>https://orcid.org/0000-0002-4281-0541</orcidid><orcidid>https://orcid.org/0000-0002-7573-8822</orcidid></search><sort><creationdate>20230404</creationdate><title>CRISPR-Cas-Driven Single Micromotor (Cas-DSM) Enables Direct Detection of Nucleic Acid Biomarkers at the Single-Molecule Level</title><author>Chen, Desheng ; Liang, Yuanwen ; Wang, Honghong ; Wang, Hui ; Su, Fengxia ; Zhang, Pengbo ; Wang, Shuhui ; Liu, Weiliang ; Li, Zhengping</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a376t-dd9cde623ad78b37f2a827b5cfcc711481b2bb2ce5244061eeaab77151fd58363</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Acids</topic><topic>Biomarkers</topic><topic>Biosensing Techniques</topic><topic>Chemistry</topic><topic>CRISPR</topic><topic>CRISPR-Cas Systems - genetics</topic><topic>DNA Primers</topic><topic>DNA probes</topic><topic>Endonuclease</topic><topic>Micromotors</topic><topic>Nucleic Acids</topic><topic>RNA</topic><topic>Target detection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Desheng</creatorcontrib><creatorcontrib>Liang, Yuanwen</creatorcontrib><creatorcontrib>Wang, Honghong</creatorcontrib><creatorcontrib>Wang, Hui</creatorcontrib><creatorcontrib>Su, Fengxia</creatorcontrib><creatorcontrib>Zhang, Pengbo</creatorcontrib><creatorcontrib>Wang, Shuhui</creatorcontrib><creatorcontrib>Liu, Weiliang</creatorcontrib><creatorcontrib>Li, Zhengping</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>Chen, Desheng</au><au>Liang, Yuanwen</au><au>Wang, Honghong</au><au>Wang, Hui</au><au>Su, Fengxia</au><au>Zhang, Pengbo</au><au>Wang, Shuhui</au><au>Liu, Weiliang</au><au>Li, Zhengping</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>CRISPR-Cas-Driven Single Micromotor (Cas-DSM) Enables Direct Detection of Nucleic Acid Biomarkers at the Single-Molecule Level</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2023-04-04</date><risdate>2023</risdate><volume>95</volume><issue>13</issue><spage>5729</spage><epage>5737</epage><pages>5729-5737</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><abstract>The target-dependent endonuclease activity (also known as the trans-cleavage activity) of CRISPR-Cas systems has stimulated great interest in the development of nascent sensing strategies for nucleic acid diagnostics. Despite many attempts, the majority of the sensitive CRISPR-Cas diagnostics strategies mainly rely on nucleic acid preamplification, which generally needs complex probes/primers designs, multiple experimental steps, and a longer testing time, as well as introducing the risk of false-positive results. In this work, we propose the CRISPR-Cas-Driven Single Micromotor (Cas-DSM), which can directly detect the nucleic acid targets at a single-molecule level with high specificity. We have demonstrated that the Cas-DSM is a reliable and practical method for the quantitative detection of DNA/RNA in various complex clinical samples as well as in individual cells without any preamplification processes. Due to the excellent features of the CRISPR/Cas system, including constant temperature, simple design, high specificity, and flexible programmability, the Cas-DSM could serve as a simple and universal platform for nucleic acid detection. More importantly, this work will provide a breakthrough for the development of next-generation amplification-free CRISPR/Cas sensing toolboxes.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>36944919</pmid><doi>10.1021/acs.analchem.2c05767</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0002-8535-5358</orcidid><orcidid>https://orcid.org/0000-0002-5116-1977</orcidid><orcidid>https://orcid.org/0000-0002-4281-0541</orcidid><orcidid>https://orcid.org/0000-0002-7573-8822</orcidid></addata></record> |
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| subjects | Acids Biomarkers Biosensing Techniques Chemistry CRISPR CRISPR-Cas Systems - genetics DNA Primers DNA probes Endonuclease Micromotors Nucleic Acids RNA Target detection |
| title | CRISPR-Cas-Driven Single Micromotor (Cas-DSM) Enables Direct Detection of Nucleic Acid Biomarkers at the Single-Molecule Level |
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