HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation
Li, Linxian1,2,3; Li, Shiyuan4; Wu, Na1,2,5; Wu, Jiacheng5; Wang, Gang3; Zhao, Guoping2,6,7; Wang, Jin8
2019-10
Source PublicationACS SYNTHETIC BIOLOGY
ISSN2161-5063
Volume8Issue:10Pages:2228-2237
DOI10.1021/acssynbio.9b00209
AbstractThe 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.
KeywordCRISPR Cas12b nucleic acid detection DNA methylation quantitation HOLMES
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Indexed BySCI
Language英语
Funding ProjectNational Natural Science Foundation of China[31922046]
WOS Research AreaBiochemistry & Molecular Biology
WOS SubjectBiochemical Research Methods
WOS IDWOS:000492115800005
PublisherAMER CHEMICAL SOC
WOS KeywordDIAGNOSTICS
Original Document TypeArticle
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Cited Times [WOS]:0   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://kms.shanghaitech.edu.cn/handle/2MSLDSTB/80513
Collection生命科学与技术学院_硕士生
Corresponding AuthorWang, Jin
Affiliation1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
2.Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, Key Lab Synthet Biol, Shanghai 200032, Peoples R China
3.Henan Univ, Sch Life Sci, State Key Lab Cotton Biol, Kaifeng 475004, Henan, Peoples R China
4.Shanghai Tolo Biotechnol Co Ltd, Shanghai 200233, Peoples R China
5.Shanghai Tech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China
6.Chinese Univ Hong Kong, Prince Wales Hosp, Dept Microbiol, Shatin, Hong Kong, Peoples R China
7.Chinese Univ Hong Kong, Prince Wales Hosp, Li KaShing Inst Hlth Sci, Shatin, Hong Kong, Peoples R China
8.Shanghai Normal Univ, Coll Life Sci, Shanghai 200234, Peoples R China
Recommended Citation
GB/T 7714
Li, Linxian,Li, Shiyuan,Wu, Na,et al. HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation[J]. ACS SYNTHETIC BIOLOGY,2019,8(10):2228-2237.
APA Li, Linxian.,Li, Shiyuan.,Wu, Na.,Wu, Jiacheng.,Wang, Gang.,...&Wang, Jin.(2019).HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation.ACS SYNTHETIC BIOLOGY,8(10),2228-2237.
MLA Li, Linxian,et al."HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation".ACS SYNTHETIC BIOLOGY 8.10(2019):2228-2237.
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