Laboratory for Genome Engineering and Synthetic Biology, Division of Biological Sciences, 4700 King Abdullah University of Science and Technology, Thuwal 23955-6900, Saudi Arabia.
ACS Synth Biol. 2020 Jun 19;9(6):1226-1233. doi: 10.1021/acssynbio.9b00507. Epub 2020 Mar 17.
For infectious diseases, rapid and accurate identification of the pathogen is critical for effective management and treatment, but diagnosis remains challenging, particularly in resource-limited areas. Methods that accurately detect pathogen nucleic acids can provide robust, accurate, rapid, and ultrasensitive technologies for point-of-care diagnosis of pathogens, and thus yield information that is invaluable for disease management and treatment. Several technologies, mostly PCR-based, have been employed for pathogen detection; however, these require expensive reagents and equipment, and skilled personnel. CRISPR/Cas systems have been used for genome editing, based on their ability to accurately recognize and cleave specific DNA and RNA sequences. Moreover, following recognition of the target sequence, certain CRISPR/Cas systems including orthologues of Cas13, Cas12a, and Cas14 exhibit collateral nonspecific catalytic activities that can be employed for nucleic acid detection, for example by degradation of a labeled nucleic acid to produce a fluorescent signal. CRISPR/Cas systems are amenable to multiplexing, thereby enabling a single diagnostic test to identify multiple targets down to attomolar (10 mol/L) concentrations of target molecules. Developing devices that couple CRISPR/Cas with lateral flow systems may allow inexpensive, accurate, highly sensitive, in-field deployable diagnostics. These sensors have myriad applications, from human health to agriculture. In this review, we discuss the recent advances in the field of CRISPR-based biosensing technologies and highlight insights of their potential use in a myriad of applications.
对于传染病而言，快速准确地鉴定病原体对于有效管理和治疗至关重要，但诊断仍然具有挑战性，尤其是在资源有限的地区。准确检测病原体核酸的方法可为即时诊断病原体提供强大、准确、快速和超灵敏的技术，从而为疾病管理和治疗提供有价值的信息。已经采用了多种技术（主要基于 PCR）来检测病原体；然而，这些技术需要昂贵的试剂和设备以及熟练的人员。CRISPR/Cas 系统基于其准确识别和切割特定 DNA 和 RNA 序列的能力，已被用于基因组编辑。此外，在识别目标序列后，某些包括 Cas13、Cas12a 和 Cas14 的同源物的 CRISPR/Cas 系统表现出旁线非特异性催化活性，可用于核酸检测，例如通过降解标记的核酸产生荧光信号。CRISPR/Cas 系统适合多重化，从而能够通过单个诊断测试识别多个目标，其检测下限可达皮摩尔（10 mol/L）浓度的目标分子。开发将 CRISPR/Cas 与横向流动系统结合的设备可能会实现廉价、准确、高灵敏度、可现场部署的诊断。这些传感器具有广泛的应用，从人类健康到农业。在这篇综述中，我们讨论了基于 CRISPR 的生物传感技术领域的最新进展，并强调了它们在众多应用中的潜在用途的见解。
