清脆的
核酸
基因组编辑
DNA
Cas9
生物
计算生物学
基因
病毒学
遗传学
作者
Li Wang,Chang Xu,Shaohui Zhang,Shang Chen,Hao Wang,Zhaojun Duan,Omar A. Al‐Hartomy,S. Wageh,Xiaosha Wen,Yi Liu,Yi Lin,Huijie Pu,Zhongjian Xie,Quan Liu,Han Zhang,Dixian Luo
出处
期刊:Applied physics reviews
[American Institute of Physics]
日期:2023-08-07
卷期号:10 (3)
被引量:5
摘要
Mpox is an infectious viral disease, and, as of January 12, 2023, about 84 560 distinct cases have been detected, involving about 110 countries since May 2022. Most clustered regularly interspaced short palindromic repeat (CRISPR)-based detection methods require long assay time because of the pre-amplification to target nucleic acids. Herein, we designed a new prototypal mpox biosensor that allows amplification-free nucleic acid detection by combining CRISPR/Cas12b system and ultrasensitive graphene field-effect transistor (gFET). The CRISPR/Cas12b–gFET enabled the detection of the mpox virus (MPXV) DNA targets at a sensitivity of ∼1 aM within about 20 min. We designed several sgRNAs to recognize the target genes of MPXV and then Cas12b protein cleaved the target genes to form a double-strand break. The specificity of sgRNAs of MPXV designed was verified among other homologous Orthopoxviruses, and the sgRNA could distinguish between recent 2022 MPXV and West African MPXV with the mutation site, showing its potential application in MPXV detection. Thus, the demonstrated CRISPR–gFET detection technology could serve as a sensitive and rapid diagnostic tool for MPXV and other DNA viruses in the future.
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