HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation
Year: 2019
Type: article
Abstract: The next-generation CRISPR-based molecular diagnostics has the merits of rapidness, accuracy, and portability. We discovered the Cas12a trans -cleavage activity against collateral single-stranded DNA (ssDNA) and employed the activity to develop a rapid nucleic acid detection system, namely HOLMES (one-hour low-cost multipurpose highly efficient system). Here, with the employment of thermophilic CRISPR-Cas12b, we create HOLMESv2 for four different applications: (1) specifically discriminating single nucleotide polymorphism (SNP); (2) simply detecting virus RNA, human cell mRNA and circular RNA; (3) conveniently quantitating target nucleic acids with a one-step system combined with LAMP amplification in a constant temperature, thus avoiding cross-contamination; (4) accurately quantitating target DNA methylation degree with the combination of Cas12b detection and bisulfite treatment. These results highlight the potential of HOLMESv2 as a promising platform for both molecular diagnostics and epigenetics applications.
Source: ACS Synthetic Biology
Authors: Linxian Li, Shiyuan Li, Na Wu, Jiacheng Wu, Gang Wang, Guoping Zhao, Jin Wang
Institutions: Henan University, Chinese Academy of Sciences, Shanghai Institutes for Biological Sciences, Center for Excellence in Molecular Plant Sciences, University of Chinese Academy of Sciences, State Key Laboratory of Cotton Biology, ShanghaiTech University, Chinese University of Hong Kong, Prince of Wales Hospital, Shanghai Normal University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: G7516945324, G8315253398
OpenAlex ID: W2974267223