دورية أكاديمية
A Rationally Designed CRISPR/Cas12a Assay Using a Multimodal Reporter for Various Readouts.
| العنوان: | A Rationally Designed CRISPR/Cas12a Assay Using a Multimodal Reporter for Various Readouts. |
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| المؤلفون: | de Dieu Habimana J; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.; University of Chinese Academy of Sciences, 19 Yuquan Road, Shijingshan District, Beijing 100049, China., Mukama O; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.; University of Chinese Academy of Sciences, 19 Yuquan Road, Shijingshan District, Beijing 100049, China.; Department of Biology, College of Science and Technology, University of Rwanda, Avenue de l'armée, Kigali 3900, Rwanda., Amissah OB; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.; University of Chinese Academy of Sciences, 19 Yuquan Road, Shijingshan District, Beijing 100049, China., Sun Y; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China., Karangwa E; Research and Development, AAFUD Industry (Zhuhai) Co. Ltd., Zhuhai 519085, China., Liu Y; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China., Mugisha S; Department of Pathology, University of California, San Diego, 9500 Gilman, La Jolla, California 92093, United States., Cheng N; Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha 410013, China., Wang L; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.; Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, China., Chen J; Department of Gynecology & Obstetrics, The Second Xiangya Hospital of Central South University, Changsha 410011, China., Deng S; Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha 410013, China., Huang R; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.; University of Chinese Academy of Sciences, 19 Yuquan Road, Shijingshan District, Beijing 100049, China.; Guangzhou Qiyuan Biomedical Co., Ltd., Guangzhou 510530, China., Li Z; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.; University of Chinese Academy of Sciences, 19 Yuquan Road, Shijingshan District, Beijing 100049, China.; Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230026, China.; GZMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou 511436, China.; GIBH-HKU Guangdong-Hong Kong Stem Cell and Regenerative Medicine Research Centre, GIBH-CUHK Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou 511436, China.; Guangzhou Qiyuan Biomedical Co., Ltd., Guangzhou 510530, China. |
| المصدر: | Analytical chemistry [Anal Chem] 2023 Aug 08; Vol. 95 (31), pp. 11741-11750. Date of Electronic Publication: 2023 Jul 28. |
| 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 , Nucleic Acids*, Biological Assay ; Octoxynol ; Nucleic Acid Amplification Techniques |
| مستخلص: | The CRISPR/Cas systems offer a programmable platform for nucleic acid detection, and CRISPR/Cas-based diagnostics (CRISPR-Dx) have demonstrated the ability to target nucleic acids with greater accuracy and flexibility. However, due to the configuration of the reporter and the underlying labeling mechanism, almost all reported CRISPR-Dx rely on a single-option readout, resulting in limitations in end-point result readouts. This is also associated with high reagent consumption and delays in diagnostic reports due to protocol differences. Herein, we report for the first time a rationally designed Cas12a-based multimodal universal reporter (CAMURE) with improved sensitivity that harnesses a dual-mode reporting system, facilitating options in end-point readouts. Through systematic configurations and optimizations, our novel universal reporter achieved a 10-fold sensitivity enhancement compared to the DETECTR reporter. Our unique and versatile reporter could be paired with various readouts, conveying the same diagnostic results. We applied our novel reporter for the detection of staphylococcal enterotoxin A due to its high implication in staphylococcal food poisoning. Integrated with loop-mediated isothermal amplification, our multimodal reporter achieved 10 CFU/mL sensitivity and excellent specificity using a real-time fluorimeter, in-tube fluorescence, and lateral flow strip readouts. We also propose, using artificially contaminated milk samples, a fast (2-5 min) Triton X-100 DNA extraction approach with a comparable yield to the commercial extraction kit. Our CAMURE could be leveraged to detect all gene-encoding SEs by simply reprogramming the guide RNA and could also be applied to the detection of other infections and disease biomarkers. |
| المشرفين على المادة: | 0 (Nucleic Acids) 9002-93-1 (Octoxynol) |
| تواريخ الأحداث: | Date Created: 20230728 Date Completed: 20230809 Latest Revision: 20230809 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1021/acs.analchem.3c01876 |
| PMID: | 37504509 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 1520-6882 |
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| DOI: | 10.1021/acs.analchem.3c01876 |