N1-Methyladenosine detection with CRISPR-Cas13a/C2c2

被引：61

作者：

Chen, Yi ^{[1
]}

Yang, Shixi ^{[1
]}

Peng, Shuang ^{[1
]}

Li, Wei ^{[1
]}

Wu, Fan ^{[1
]}

Yao, Qian ^{[1
]}

Wang, Fang ^{[2
]}

Weng, Xiaocheng ^{[1
]}

Zhou, Xiang ^{[1
]}

机构：

[1] Wuhan Univ, Coll Chem & Mol Sci, Key Lab Biomed Polymers,Inst Adv Studies, Hubei Prov Key Lab Allergy & Immunol,Minist Educ, Wuhan 430072, Hubei, Peoples R China

[2] Wuhan Univ, Sch Pharmaceut Sci, Wuhan 430071, Hubei, Peoples R China

来源：

CHEMICAL SCIENCE | 2019年 / 10卷 / 10期

基金：

中国国家自然科学基金;

关键词：

RNA; METHYLTRANSFERASE; N-1-METHYLADENOSINE; METHYLATION; SEQUENCE; ENDORIBONUCLEASE; IDENTIFICATION; BIOGENESIS; METHYLOME;

D O I：

10.1039/c8sc03408g

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Recent studies suggested that the widespread presence of N-1-methyladenosine (m(1)A) plays a very important role in environmental stress, ribosome biogenesis and antibiotic resistance. The RNA-guided, RNA-targeting CRISPR Cas13a exhibits a "collateral effect" of promiscuous RNase activity upon target recognition with high sensitivity. Inspired by the advantage of CRISPR Cas13a, we designed a system to detect m(1)A induced mismatch, providing a rapid, simple and fluorescence-based m(1)A detection. For A-ssRNA, the Cas13a-based molecular detection platform showed a high fluorescence signal. For m(1)A-ssRNA, there is an about 90% decline of fluorescence. Moreover, this approach can also be used to quantify m(1)A in RNAs and applied for the analysis of dynamic m(1)A demethylation of 28S rRNA with AlkB.

引用

页码：2975 / 2979

页数：5

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