Signal-On Fluorescence Biosensor for Highly Sensitive Detection of miRNA-21 Based on DNAzyme Assisted Double-Hairpin Molecular Beacon
Although miRNAs exist in small quantities in the human body, they are closely related to the abnormal expression of genes in diseases such as tumors. Therefore, sensitive detection of miRNAs is very important for the prevention and treatment of various tumors and major diseases. The purpose of this...
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MDPI AG
2022-04-01
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| Online Access: | https://www.mdpi.com/2079-6374/12/5/276 |
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| author | Chenxin Fang Yuxing Yang Shuhao Zou Ping Ouyang Yang Qing Jialun Han Haiyu Li Zhencui Wang Jie Du |
| author_facet | Chenxin Fang Yuxing Yang Shuhao Zou Ping Ouyang Yang Qing Jialun Han Haiyu Li Zhencui Wang Jie Du |
| author_sort | Chenxin Fang |
| collection | DOAJ |
| description | Although miRNAs exist in small quantities in the human body, they are closely related to the abnormal expression of genes in diseases such as tumors. Therefore, sensitive detection of miRNAs is very important for the prevention and treatment of various tumors and major diseases. The purpose of this study is to develop a label-free sensing strategy based on the co-action of double-hairpin molecular beacons and deoxyribozymes (DNAzymes) for highly sensitive detection of miRNA-21. The target miRNA-21 promotes the assembly of DNAzyme with a complete catalytic core region. At the presence of Mg<sup>2+</sup>, DNAzyme cuts a substrate into short chains, which open the double hairpin molecular beacon, and then form G-quadruplexs at both ends, specifically binding more ThT to generate a amplified fluorescent signal. The cut substrate will be replaced by the uncut ones in the next stage, increasing the concentration of reactants, and thus further improving the fluorescence intensity. This DNAzyme assisted double hairpin molecular beacon has a certain degree of discrimination for substances with single base mismatches, and the detection limit of miRNA-21 is 0.13 pM, lower than that of the many other analysis. Further, this detection has good selectivity and sensitivity in serum. Therefore, this strategy provides a simple, fast and low-cost platform for the sensitive detection of miRNA-21, having potential applications in early cancer diagnosis. |
| first_indexed | 2024-03-10T03:15:26Z |
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| id | doaj.art-4e73f3ff4c71407a8fefed9c6f857b4f |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-10T03:15:26Z |
| publishDate | 2022-04-01 |
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| series | Biosensors |
| spelling | doaj.art-4e73f3ff4c71407a8fefed9c6f857b4f2023-11-23T10:15:10ZengMDPI AGBiosensors2079-63742022-04-0112527610.3390/bios12050276Signal-On Fluorescence Biosensor for Highly Sensitive Detection of miRNA-21 Based on DNAzyme Assisted Double-Hairpin Molecular BeaconChenxin Fang0Yuxing Yang1Shuhao Zou2Ping Ouyang3Yang Qing4Jialun Han5Haiyu Li6Zhencui Wang7Jie Du8College of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaCollege of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaCollege of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaCollege of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaCollege of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaCollege of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaCollege of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaCollege of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaCollege of Materials Science and Engineering, Hainan University, Haikou 570228, ChinaAlthough miRNAs exist in small quantities in the human body, they are closely related to the abnormal expression of genes in diseases such as tumors. Therefore, sensitive detection of miRNAs is very important for the prevention and treatment of various tumors and major diseases. The purpose of this study is to develop a label-free sensing strategy based on the co-action of double-hairpin molecular beacons and deoxyribozymes (DNAzymes) for highly sensitive detection of miRNA-21. The target miRNA-21 promotes the assembly of DNAzyme with a complete catalytic core region. At the presence of Mg<sup>2+</sup>, DNAzyme cuts a substrate into short chains, which open the double hairpin molecular beacon, and then form G-quadruplexs at both ends, specifically binding more ThT to generate a amplified fluorescent signal. The cut substrate will be replaced by the uncut ones in the next stage, increasing the concentration of reactants, and thus further improving the fluorescence intensity. This DNAzyme assisted double hairpin molecular beacon has a certain degree of discrimination for substances with single base mismatches, and the detection limit of miRNA-21 is 0.13 pM, lower than that of the many other analysis. Further, this detection has good selectivity and sensitivity in serum. Therefore, this strategy provides a simple, fast and low-cost platform for the sensitive detection of miRNA-21, having potential applications in early cancer diagnosis.https://www.mdpi.com/2079-6374/12/5/276signal amplificationfluorescent biosensorDNAzymemiRNA detectionhigh sensitivity |
| spellingShingle | Chenxin Fang Yuxing Yang Shuhao Zou Ping Ouyang Yang Qing Jialun Han Haiyu Li Zhencui Wang Jie Du Signal-On Fluorescence Biosensor for Highly Sensitive Detection of miRNA-21 Based on DNAzyme Assisted Double-Hairpin Molecular Beacon Biosensors signal amplification fluorescent biosensor DNAzyme miRNA detection high sensitivity |
| title | Signal-On Fluorescence Biosensor for Highly Sensitive Detection of miRNA-21 Based on DNAzyme Assisted Double-Hairpin Molecular Beacon |
| title_full | Signal-On Fluorescence Biosensor for Highly Sensitive Detection of miRNA-21 Based on DNAzyme Assisted Double-Hairpin Molecular Beacon |
| title_fullStr | Signal-On Fluorescence Biosensor for Highly Sensitive Detection of miRNA-21 Based on DNAzyme Assisted Double-Hairpin Molecular Beacon |
| title_full_unstemmed | Signal-On Fluorescence Biosensor for Highly Sensitive Detection of miRNA-21 Based on DNAzyme Assisted Double-Hairpin Molecular Beacon |
| title_short | Signal-On Fluorescence Biosensor for Highly Sensitive Detection of miRNA-21 Based on DNAzyme Assisted Double-Hairpin Molecular Beacon |
| title_sort | signal on fluorescence biosensor for highly sensitive detection of mirna 21 based on dnazyme assisted double hairpin molecular beacon |
| topic | signal amplification fluorescent biosensor DNAzyme miRNA detection high sensitivity |
| url | https://www.mdpi.com/2079-6374/12/5/276 |
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