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HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation | |
2019-10 | |
Source Publication | ACS SYNTHETIC BIOLOGY
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ISSN | 2161-5063 |
Volume | 8Issue:10Pages:2228-2237 |
Status | 已发表 |
DOI | 10.1021/acssynbio.9b00209 |
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. |
Keyword | CRISPR Cas12b nucleic acid detection DNA methylation quantitation HOLMES |
URL | 查看原文 |
Indexed By | SCI ; SCIE |
Language | 英语 |
Funding Project | National Natural Science Foundation of China[31922046] |
WOS Research Area | Biochemistry & Molecular Biology |
WOS Subject | Biochemical Research Methods |
WOS ID | WOS:000492115800005 |
Publisher | AMER CHEMICAL SOC |
WOS Keyword | DIAGNOSTICS |
Original Document Type | Article |
Citation statistics | |
Document Type | 期刊论文 |
Identifier | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/80513 |
Collection | 生命科学与技术学院_硕士生 |
Corresponding Author | Wang, Jin |
Affiliation | 1.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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