Towards application of CRISPR-Cas12a in the design of modern viral DNA detection tools (Review)

被引：99
作者：
Dronina, Julija [1 ]
Samukaite-Bubniene, Urte [1 ,2 ]
Ramanavicius, Arunas [1 ,2 ]
机构：
[1] Ctr Phys Sci & Technol, Dept Funct Mat & Elect, Lab Nanotechnol, Sauletekio Av 3, Vilnius, Lithuania
[2] Vilnius Univ, Fac Chem & Geosci, Dept Phys Chem, Naugarduko Str 24, LT-03225 Vilnius, Lithuania
来源：
JOURNAL OF NANOBIOTECHNOLOGY | 2022年 / 20卷 / 01期
关键词：
COVID-19; CRISPR-Cas; DNA-biosensors; SARS-CoV-2; virus; CRISPR-Cas12a; Bioanalytical system; CRISPR-CAS SYSTEMS; RNA-GUIDED ENDONUCLEASE; CRYSTAL-STRUCTURE; EVOLUTIONARY CLASSIFICATION; STRUCTURAL BASIS; VIRUS STRUCTURE; BINDING DOMAIN; COAT PROTEIN; CPF1; GENE;
D O I：
10.1186/s12951-022-01246-7
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Early detection of viral pathogens by DNA-sensors in clinical samples, contaminated foods, soil or water can dramatically improve clinical outcomes and reduce the socioeconomic impact of diseases such as COVID-19. Clustered regularly interspaced short palindromic repeat (CRISPR) and its associated protein Cas12a (previously known as CRISPR-Cpf1) technology is an innovative new-generation genomic engineering tool, also known as 'genetic scissors', that has demonstrated the accuracy and has recently been effectively applied as appropriate (E-CRISPR) DNA-sensor to detect the nucleic acid of interest. The CRISPR-Cas12a from Prevotella and Francisella 1 are guided by a short CRISPR RNA (gRNA). The unique simultaneous cis- and trans- DNA cleavage after target sequence recognition at the PAM site, sticky-end (5-7 bp) employment, and ssDNA/dsDNA hybrid cleavage strategies to manipulate the attractive nature of CRISPR-Cas12a are reviewed. DNA-sensors based on the CRISPR-Cas12a technology for rapid, robust, sensitive, inexpensive, and selective detection of virus DNA without additional sample purification, amplification, fluorescent-agent- and/or quencher-labeling are relevant and becoming increasingly important in industrial and medical applications. In addition, CRISPR-Cas12a system shows great potential in the field of E-CRISPR-based bioassay research technologies. Therefore, we are highlighting insights in this research direction.
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页数：15
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