CDetection: CRISPR-Cas12b-based DNA detection with sub-attomolar sensitivity and single-base specificity

被引:243
|
作者
Teng, Fei [1 ,2 ,3 ]
Guo, Lu [1 ,2 ,3 ]
Cui, Tongtong [1 ,2 ,3 ]
Wang, Xin-Ge [1 ,2 ,3 ]
Xu, Kai [1 ,2 ,3 ]
Gao, Qingqin [1 ,2 ,3 ]
Zhou, Qi [1 ,2 ,3 ]
Li, Wei [1 ,2 ,3 ]
机构
[1] Chinese Acad Sci, State Key Lab Stem Cell & Reprod Biol, Inst Zool, Beijing 100101, Peoples R China
[2] Chinese Acad Sci, Inst Stem Cell & Regenerat, Beijing 100101, Peoples R China
[3] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
基金
中国国家自然科学基金;
关键词
RNA-GUIDED ENDONUCLEASE; NUCLEIC-ACID DETECTION; CPF1; NUCLEASES;
D O I
10.1186/s13059-019-1742-z
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
CRISPR-based nucleic acid detection methods are reported to facilitate rapid and sensitive DNA detection. However, precise DNA detection at the single-base resolution and its wide applications including high-fidelity SNP genotyping remain to be explored. Here we develop a Cas12b-mediated DNA detection (CDetection) strategy, which shows higher sensitivity on examined targets compared with the previously reported Cas12a-based detection platform. Moreover, we show that CDetection can distinguish differences at the single-base level upon combining the optimized tuned guide RNA (tgRNA). Therefore, our findings highlight the high sensitivity and accuracy of CDetection, which provides an efficient and highly practical platform for DNA detection.
引用
收藏
页数:7
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