دورية أكاديمية
Allosteric Activator-Regulated CRISPR/Cas12a System Enables Biosensing and Imaging of Intracellular Endogenous and Exogenous Targets.
| العنوان: | Allosteric Activator-Regulated CRISPR/Cas12a System Enables Biosensing and Imaging of Intracellular Endogenous and Exogenous Targets. |
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| المؤلفون: | Li QN; State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina., Ma AX; State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina., Wang DX; State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina., Dai ZQ; State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina., Wu SL; State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina., Lu S; State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina., Zhu LN; Department of Chemistry, School of Science, Tianjin University, Tianjin, 300354, PRChina., Jiang HX; Agro-Environmental Protection Institute, Key Laboratory for Environmental Factors Control of Agro-product Quality Safety, Laboratory of Environmental Factors Risk Assessment of Agro-Product Quality Safety, Ministry of Agriculture, Tianjin, 300191, PRChina., Pang DW; State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina., Kong DM; State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, Research Centre for Analytical Sciences, College of Chemistry, Nankai University, Tianjin, 300071, PRChina. |
| المصدر: | Analytical chemistry [Anal Chem] 2024 Apr 23; Vol. 96 (16), pp. 6426-6435. Date of Electronic Publication: 2024 Apr 11. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 0370536 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-6882 (Electronic) Linking ISSN: 00032700 NLM ISO Abbreviation: Anal Chem Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, American Chemical Society. |
| مواضيع طبية MeSH: | CRISPR-Cas Systems*/genetics , Biosensing Techniques*/methods , MicroRNAs*/analysis , MicroRNAs*/metabolism, Humans ; Allosteric Regulation ; CRISPR-Associated Proteins/metabolism ; Endodeoxyribonucleases/metabolism ; Endodeoxyribonucleases/chemistry ; Bacterial Proteins/metabolism ; Bacterial Proteins/chemistry ; Bacterial Proteins/genetics ; HEK293 Cells |
| مستخلص: | Sensors designed based on the trans -cleavage activity of CRISPR/Cas12a systems have opened up a new era in the field of biosensing. The current design of CRISPR/Cas12-based sensors in the "on-off-on" mode mainly focuses on programming the activator strand (AS) to indirectly switch the trans -cleavage activity of Cas12a in response to target information. However, this design usually requires the help of additional auxiliary probes to keep the activator strand in an initially "blocked" state. The length design and dosage of the auxiliary probe need to be strictly optimized to ensure the lowest background and the best signal-to-noise ratio. This will inevitably increase the experiment complexity. To solve this problem, we propose using AS after the "RESET" effect to directly regulate the Cas12a enzymatic activity. Initially, the activator strand was rationally designed to be embedded in a hairpin structure to deprive its ability to activate the CRISPR/Cas12a system. When the target is present, target-mediated strand displacement causes the conformation change in the AS, the hairpin structure is opened, and the CRISPR/Cas12a system is reactivated; the switchable structure of AS can be used to regulate the degree of activation of Cas12a according to the target concentration. Due to the advantages of low background and stability, the CRISPR/Cas12a-based strategy can not only image endogenous biomarkers (miR-21) in living cells but also enable long-term and accurate imaging analysis of the process of exogenous virus invasion of cells. Release and replication of virus genome in host cells are indispensable hallmark events of cell infection by virus; sensitive monitoring of them is of great significance to revealing virus infection mechanism and defending against viral diseases. |
| المشرفين على المادة: | 0 (MicroRNAs) 0 (CRISPR-Associated Proteins) 0 (MIRN21 microRNA, human) EC 3.1.- (Cas12a protein) EC 3.1.- (Endodeoxyribonucleases) 0 (Bacterial Proteins) |
| تواريخ الأحداث: | Date Created: 20240411 Date Completed: 20240424 Latest Revision: 20240429 |
| رمز التحديث: | 20240429 |
| DOI: | 10.1021/acs.analchem.4c00555 |
| PMID: | 38604773 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 1520-6882 |
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| DOI: | 10.1021/acs.analchem.4c00555 |