{"title":"利用 CRISPR/Cas 系统检测 DNA 和 RNA:原理、技术和挑战。","authors":"Heyjin Son","doi":"10.3390/bios14100460","DOIUrl":null,"url":null,"abstract":"<p><p>The emergence of CRISPR/Cas systems has revolutionized the field of molecular diagnostics with their high specificity and sensitivity. This review provides a comprehensive overview of the principles and recent advancements in harnessing CRISPR/Cas systems for detecting DNA and RNA. Beginning with an exploration of the molecular mechanisms of key Cas proteins underpinning CRISPR/Cas systems, the review navigates the detection of both pathogenic and non-pathogenic nucleic acids, emphasizing the pivotal role of CRISPR in identifying diverse genetic materials. The discussion extends to the integration of CRISPR/Cas systems with various signal-readout techniques, including fluorescence, electrochemical, and colorimetric, as well as imaging and biosensing methods, highlighting their advantages and limitations in practical applications. Furthermore, a critical analysis of challenges in the field, such as target amplification, multiplexing, and quantitative detection, underscores areas requiring further refinement. Finally, the review concludes with insights into the future directions of CRISPR-based nucleic acid detection, emphasizing the potential of these systems to continue driving innovation in diagnostics, with broad implications for research, clinical practice, and biotechnology.</p>","PeriodicalId":48608,"journal":{"name":"Biosensors-Basel","volume":null,"pages":null},"PeriodicalIF":4.9000,"publicationDate":"2024-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506544/pdf/","citationCount":"0","resultStr":"{\"title\":\"Harnessing CRISPR/Cas Systems for DNA and RNA Detection: Principles, Techniques, and Challenges.\",\"authors\":\"Heyjin Son\",\"doi\":\"10.3390/bios14100460\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The emergence of CRISPR/Cas systems has revolutionized the field of molecular diagnostics with their high specificity and sensitivity. This review provides a comprehensive overview of the principles and recent advancements in harnessing CRISPR/Cas systems for detecting DNA and RNA. Beginning with an exploration of the molecular mechanisms of key Cas proteins underpinning CRISPR/Cas systems, the review navigates the detection of both pathogenic and non-pathogenic nucleic acids, emphasizing the pivotal role of CRISPR in identifying diverse genetic materials. The discussion extends to the integration of CRISPR/Cas systems with various signal-readout techniques, including fluorescence, electrochemical, and colorimetric, as well as imaging and biosensing methods, highlighting their advantages and limitations in practical applications. Furthermore, a critical analysis of challenges in the field, such as target amplification, multiplexing, and quantitative detection, underscores areas requiring further refinement. Finally, the review concludes with insights into the future directions of CRISPR-based nucleic acid detection, emphasizing the potential of these systems to continue driving innovation in diagnostics, with broad implications for research, clinical practice, and biotechnology.</p>\",\"PeriodicalId\":48608,\"journal\":{\"name\":\"Biosensors-Basel\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.9000,\"publicationDate\":\"2024-09-26\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11506544/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biosensors-Basel\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.3390/bios14100460\",\"RegionNum\":3,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biosensors-Basel","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.3390/bios14100460","RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0
摘要
CRISPR/Cas 系统以其高特异性和高灵敏度彻底改变了分子诊断领域。本综述全面概述了利用 CRISPR/Cas 系统检测 DNA 和 RNA 的原理和最新进展。本综述从探讨支持 CRISPR/Cas 系统的关键 Cas 蛋白的分子机制开始,探讨了病原体和非病原体核酸的检测,强调了 CRISPR 在识别各种遗传物质方面的关键作用。讨论延伸到 CRISPR/Cas 系统与各种信号读出技术(包括荧光、电化学和比色法)以及成像和生物传感方法的整合,强调了它们在实际应用中的优势和局限性。此外,还对目标放大、多路复用和定量检测等领域的挑战进行了批判性分析,强调了需要进一步完善的领域。最后,综述对基于 CRISPR 的核酸检测的未来发展方向进行了深入分析,强调了这些系统继续推动诊断创新的潜力,对研究、临床实践和生物技术具有广泛的影响。
Harnessing CRISPR/Cas Systems for DNA and RNA Detection: Principles, Techniques, and Challenges.
The emergence of CRISPR/Cas systems has revolutionized the field of molecular diagnostics with their high specificity and sensitivity. This review provides a comprehensive overview of the principles and recent advancements in harnessing CRISPR/Cas systems for detecting DNA and RNA. Beginning with an exploration of the molecular mechanisms of key Cas proteins underpinning CRISPR/Cas systems, the review navigates the detection of both pathogenic and non-pathogenic nucleic acids, emphasizing the pivotal role of CRISPR in identifying diverse genetic materials. The discussion extends to the integration of CRISPR/Cas systems with various signal-readout techniques, including fluorescence, electrochemical, and colorimetric, as well as imaging and biosensing methods, highlighting their advantages and limitations in practical applications. Furthermore, a critical analysis of challenges in the field, such as target amplification, multiplexing, and quantitative detection, underscores areas requiring further refinement. Finally, the review concludes with insights into the future directions of CRISPR-based nucleic acid detection, emphasizing the potential of these systems to continue driving innovation in diagnostics, with broad implications for research, clinical practice, and biotechnology.
Biosensors-BaselBiochemistry, Genetics and Molecular Biology-Clinical Biochemistry
CiteScore
6.60
自引率
14.80%
发文量
983
审稿时长
11 weeks
期刊介绍:
Biosensors (ISSN 2079-6374) provides an advanced forum for studies related to the science and technology of biosensors and biosensing. It publishes original research papers, comprehensive reviews and communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files and software regarding the full details of the calculation or experimental procedure, if unable to be published in a normal way, can be deposited as supplementary electronic material.