HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation

doi:10.1021/acssynbio.9b00209

	HOLMESv2: A CRISPR-Cas12b-Assisted Platform for Nucleic Acid Detection and DNA Methylation Quantitation
	Li, Linxian 1,2,3; Li, Shiyuan 4; Wu, Na1,2,5 ; Wu, Jiacheng5 ; Wang, Gang 3; Zhao, Guoping 2,6,7; Wang, Jin 8
	2019-10
发表期刊	ACS SYNTHETIC BIOLOGY
ISSN	2161-5063
卷号	8 期号:10 页码:2228-2237
发表状态	已发表
DOI	10.1021/acssynbio.9b00209
摘要	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.
关键词	CRISPR Cas12b nucleic acid detection DNA methylation quantitation HOLMES
URL	查看原文
收录类别	SCI ; SCIE
语种	英语
资助项目	National Natural Science Foundation of China[31922046]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemical Research Methods
WOS记录号	WOS:000492115800005
出版者	AMER CHEMICAL SOC
WOS关键词	DIAGNOSTICS
原始文献类型	Article
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/80513
专题	生命科学与技术学院_硕士生
通讯作者	Wang, Jin
作者单位	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
推荐引用方式 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.