An RNA-based catalytic hairpin assembly circuit coupled with CRISPR-Cas12a for one-step detection of microRNAs.
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| Title: | An RNA-based catalytic hairpin assembly circuit coupled with CRISPR-Cas12a for one-step detection of microRNAs. |
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| Authors: | Chen, Pinru1 (AUTHOR), Wang, Luying1 (AUTHOR), Qin, Peipei1 (AUTHOR), Yin, Bin-Cheng1,2,3 (AUTHOR) binchengyin@ecust.edu.cn, Ye, Bang-Ce1,2 (AUTHOR) bcye@ecust.edu.cn |
| Source: | Biosensors & Bioelectronics. Jul2022, Vol. 207, pN.PAG-N.PAG. 1p. |
| Subjects: | Hairpin (Genetics), MicroRNA, Detection limit, Molecular diagnosis, Cell lines, Nucleic acids, CRISPRs |
| Abstract: | CRISPR-Cas nuclease-based nucleic acid detection has exhibited extraordinary value in the field of molecular diagnostics, but it usually involves two separate reaction steps of nucleic acid amplification and Cas-based endpoint detection, resulting in the use of multiple enzymes, inconvenient operation, and potential carry-over contamination. Here, we propose an RNA-based catalytic hairpin assembly (CHA) circuit coupled with CRISPR-Cas12a for one-step detection of microRNAs (miRNAs) at an isothermal condition. This method relies on the rational design of a spacer-blocking crRNA as a bridge between the two systems. The target miRNA can specifically trigger RNA-based CHA and induce a configurational change of the blocked crRNAs into precursor crRNAs (pre-crRNAs), which can be processed into mature crRNAs to function by leveraging the inherent RNase activities of Cas12a. In this way, the developed circuit achieves a femtomolar detection limit and shows an accurate detection of miRNA levels in different cell lines. Therefore, our method would provide a new paradigm to develop miRNA detection methods based on the CRISPR/Cas system. • A method for one-step microRNA detection based on CRISPR-Cas12a-assisted catalytic hairpin assembly (CHA) was developed. • The RNase and DNase activities of Cas12a were fully exploited. • It integrated the inherently high sensitivity and specificity both from CHA and CRISPR-Cas12a system. • This method was able to detect target miRNA down to femtomolar concentration. [ABSTRACT FROM AUTHOR] |
| Copyright of Biosensors & Bioelectronics is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
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| Header | DbId: egs DbLabel: Engineering Source An: 156471162 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: An RNA-based catalytic hairpin assembly circuit coupled with CRISPR-Cas12a for one-step detection of microRNAs. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Chen%2C+Pinru%22">Chen, Pinru</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Wang%2C+Luying%22">Wang, Luying</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Qin%2C+Peipei%22">Qin, Peipei</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Yin%2C+Bin-Cheng%22">Yin, Bin-Cheng</searchLink><relatesTo>1,2,3</relatesTo> (AUTHOR)<i> binchengyin@ecust.edu.cn</i><br /><searchLink fieldCode="AR" term="%22Ye%2C+Bang-Ce%22">Ye, Bang-Ce</searchLink><relatesTo>1,2</relatesTo> (AUTHOR)<i> bcye@ecust.edu.cn</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Biosensors+%26+Bioelectronics%22">Biosensors & Bioelectronics</searchLink>. Jul2022, Vol. 207, pN.PAG-N.PAG. 1p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Hairpin+%28Genetics%29%22">Hairpin (Genetics)</searchLink><br /><searchLink fieldCode="DE" term="%22MicroRNA%22">MicroRNA</searchLink><br /><searchLink fieldCode="DE" term="%22Detection+limit%22">Detection limit</searchLink><br /><searchLink fieldCode="DE" term="%22Molecular+diagnosis%22">Molecular diagnosis</searchLink><br /><searchLink fieldCode="DE" term="%22Cell+lines%22">Cell lines</searchLink><br /><searchLink fieldCode="DE" term="%22Nucleic+acids%22">Nucleic acids</searchLink><br /><searchLink fieldCode="DE" term="%22CRISPRs%22">CRISPRs</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: CRISPR-Cas nuclease-based nucleic acid detection has exhibited extraordinary value in the field of molecular diagnostics, but it usually involves two separate reaction steps of nucleic acid amplification and Cas-based endpoint detection, resulting in the use of multiple enzymes, inconvenient operation, and potential carry-over contamination. Here, we propose an RNA-based catalytic hairpin assembly (CHA) circuit coupled with CRISPR-Cas12a for one-step detection of microRNAs (miRNAs) at an isothermal condition. This method relies on the rational design of a spacer-blocking crRNA as a bridge between the two systems. The target miRNA can specifically trigger RNA-based CHA and induce a configurational change of the blocked crRNAs into precursor crRNAs (pre-crRNAs), which can be processed into mature crRNAs to function by leveraging the inherent RNase activities of Cas12a. In this way, the developed circuit achieves a femtomolar detection limit and shows an accurate detection of miRNA levels in different cell lines. Therefore, our method would provide a new paradigm to develop miRNA detection methods based on the CRISPR/Cas system. • A method for one-step microRNA detection based on CRISPR-Cas12a-assisted catalytic hairpin assembly (CHA) was developed. • The RNase and DNase activities of Cas12a were fully exploited. • It integrated the inherently high sensitivity and specificity both from CHA and CRISPR-Cas12a system. • This method was able to detect target miRNA down to femtomolar concentration. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Biosensors & Bioelectronics is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1016/j.bios.2022.114152 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 1 StartPage: N.PAG Subjects: – SubjectFull: Hairpin (Genetics) Type: general – SubjectFull: MicroRNA Type: general – SubjectFull: Detection limit Type: general – SubjectFull: Molecular diagnosis Type: general – SubjectFull: Cell lines Type: general – SubjectFull: Nucleic acids Type: general – SubjectFull: CRISPRs Type: general Titles: – TitleFull: An RNA-based catalytic hairpin assembly circuit coupled with CRISPR-Cas12a for one-step detection of microRNAs. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Chen, Pinru – PersonEntity: Name: NameFull: Wang, Luying – PersonEntity: Name: NameFull: Qin, Peipei – PersonEntity: Name: NameFull: Yin, Bin-Cheng – PersonEntity: Name: NameFull: Ye, Bang-Ce IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 07 Text: Jul2022 Type: published Y: 2022 Identifiers: – Type: issn-print Value: 09565663 Numbering: – Type: volume Value: 207 Titles: – TitleFull: Biosensors & Bioelectronics Type: main |
| ResultId | 1 |
