Biophotonics Group, Quant Lase Imaging Laboratory, Quant Lase Lab LLC, Abu Dhabi, United Arab Emirates.
Mol Med Rep. 2023 Jun;27(6). doi: 10.3892/mmr.2023.13005. Epub 2023 May 5.
Sudden viral outbreaks have increased in the early part of the 21st century, such as those of severe acute respiratory syndrome coronavirus (SARS‑CoV), Middle East respiratory syndrome corona virus, and SARS‑CoV‑2, owing to increased human access to wildlife habitats. Therefore, the likelihood of zoonotic transmission of human‑associated viruses has increased. The emergence of severe acute respiratory syndrome coronavirus 2 in China and its spread worldwide within months have highlighted the need to be ready with advanced diagnostic and antiviral approaches to treat newly emerging diseases with minimal harm to human health. The gold‑standard molecular diagnostic approaches currently used are time‑consuming, require trained personnel and sophisticated equipment, and therefore cannot be used as point‑of‑care devices for widespread monitoring and surveillance. Clustered regularly interspaced short palindromic repeats (CRISPR)‑associated (Cas) systems are widespread and have been reported in bacteria, archaea and bacteriophages. CRISPR‑Cas systems are organized into CRISPR arrays and adjacent Cas proteins. The detection and in‑depth biochemical characterization of class 2 type V and VI CRISPR‑Cas systems and orthologous proteins such as Cas12 and Cas13 have led to the development of CRISPR‑based diagnostic approaches, which have been used to detect viral diseases and distinguish between serotypes and subtypes. CRISPR‑based diagnostic approaches detect human single nucleotide polymorphisms in samples from patients with cancer and are used as antiviral agents to detect and destroy viruses that contain RNA as a genome. CRISPR‑based diagnostic approaches are likely to improve disease detection methods in the 21st century owing to their ease of development, low cost, reduced turnaround time, multiplexing and ease of deployment. The present review discusses the biochemical properties of Cas12 and Cas13 orthologs in viral disease detection and other applications. The present review expands the scope of CRISPR‑based diagnostic approaches to detect diseases and fight viruses as antivirals.
21 世纪初,由于人类更多地接触野生动物栖息地,突发病毒性疫情有所增加,例如严重急性呼吸系统综合征冠状病毒(SARS-CoV)、中东呼吸系统综合征冠状病毒和 SARS-CoV-2。因此,人类相关病毒发生人畜共患传播的可能性增加。SARS-CoV-2 在中国的出现及其在数月内在全球范围内的传播,突显了有必要准备先进的诊断和抗病毒方法,以在对人类健康的危害最小的情况下治疗新出现的疾病。目前使用的金标准分子诊断方法耗时,需要经过培训的人员和复杂的设备,因此不能用作广泛监测和监视的即时诊断设备。规律成簇间隔短回文重复(CRISPR)相关(Cas)系统广泛存在于细菌、古菌和噬菌体中。CRISPR-Cas 系统被组织成 CRISPR 阵列和相邻的 Cas 蛋白。对 2 类 5 型和 6 型 CRISPR-Cas 系统以及同源蛋白(如 Cas12 和 Cas13)的检测和深入生化特性分析,导致了基于 CRISPR 的诊断方法的发展,这些方法已被用于检测病毒性疾病,并区分血清型和亚型。基于 CRISPR 的诊断方法可检测癌症患者样本中的人类单核苷酸多态性,并用作抗病毒药物来检测和破坏含有 RNA 基因组的病毒。由于易于开发、成本低、周转时间短、多重检测和易于部署,基于 CRISPR 的诊断方法有望改善 21 世纪的疾病检测方法。本文讨论了 Cas12 和 Cas13 同源物在病毒疾病检测和其他应用中的生化特性。本文扩展了基于 CRISPR 的诊断方法的范围,以作为抗病毒药物来检测疾病和对抗病毒。