Infectious Diseases and Immunology Division, CSIR-Indian Institute of Chemical Biology, Kolkata, India.
Department of Botany, Visva-Bharati, Bolpur, India.
Appl Microbiol Biotechnol. 2021 Oct;105(20):7593-7605. doi: 10.1007/s00253-021-11583-6. Epub 2021 Sep 20.
The emergence of SARS-CoV-2 has brought the world to a standstill, and till date, effective treatments and diagnostics against this idiosyncratic pathogen are lacking. As compared to the standard WHO/CDC qPCR detection method, which consumes several hours for detection, CRISPR-based SHERLOCK, DETECTR, and FELUDA have emerged as rapid diagnostic tools for the detection of the RNA genome of SARS-CoV-2 within an hour with 100% accuracy, specificity, and sensitivity. These attributes of CRISPR-based detection technologies have taken themselves one step ahead of available detection systems and are emerging as an inevitable tool for quick detection of the virus. Further, the discovery of Cas13s nucleases and their orthologs has opened a new corridor for exploitation of Cas13s as an antiviral therapy against SARS-CoV-2 and other viral diseases. One such approach is Prophylactic Antiviral CRISPR in huMAN cells (PACMAN), which needs a long haul to bring into therapy. The approval of SHERLOCK as the first CRISPR-based SARS-CoV-2 test kit by the FDA, for emergency diagnosis of COVID-19 patients, has given positive hope to scientists that sooner human trials of CRISPR-based therapy will be ratified. In this review, we have extensively reviewed the present CRISPR-based approaches, challenges, and future prospects in the light of diagnostics and therapeutics against SARS-CoV-2. KEY POINTS: • The discovery of Cas12 and Cas13 siblings allowed scientists to detect the viral genes. • Cas13d's identification aided scientists in precisely cleaving the SARS-CoV-2 ssRNA. • CRISPR-Cas system acts as "molecular detector and antiviral proctor."
SARS-CoV-2 的出现使世界陷入停顿，迄今为止，针对这种特殊病原体还缺乏有效的治疗和诊断方法。与标准的世卫组织/CDC qPCR 检测方法相比，该方法需要几个小时进行检测，而基于 CRISPR 的 SHERLOCK、DETECTR 和 FELUDA 已成为快速诊断工具，可在一小时内以 100%的准确性、特异性和灵敏度检测 SARS-CoV-2 的 RNA 基因组。基于 CRISPR 的检测技术的这些特性使其比现有检测系统更先进一步，并且正在成为快速检测病毒的必不可少的工具。此外，Cas13 核酸酶及其同源物的发现为 Cas13 作为针对 SARS-CoV-2 和其他病毒疾病的抗病毒疗法的开发开辟了新的途径。一种这样的方法是 Prophylactic Antiviral CRISPR in huMAN cells (PACMAN)，需要很长时间才能投入治疗。FDA 批准 SHERLOCK 作为首个基于 CRISPR 的 SARS-CoV-2 检测试剂盒，用于 COVID-19 患者的紧急诊断，这给科学家们带来了积极的希望，即基于 CRISPR 的治疗的人体试验将很快获得批准。在这篇综述中，我们根据针对 SARS-CoV-2 的诊断和治疗，广泛回顾了基于 CRISPR 的方法、挑战和未来前景。关键点：• Cas12 和 Cas13 兄弟姐妹的发现使科学家能够检测病毒基因。• Cas13d 的鉴定帮助科学家精确地切割 SARS-CoV-2 的 ssRNA。• CRISPR-Cas 系统充当“分子探测器和抗病毒监督者”。
