Detection of target DNA with a visual CRISPR-associated hyperbranched rolling circle amplification technique

被引：6

作者：

Zhang, Beibei

Li, Miao

Wei, Yuanmeng

Wang, Jiaojiao

Wang, Yange

Shi, Pingling

Tang, He

Song, Zongming ^{[1
]}

机构：

[1] Zhengzhou Univ, Henan Univ, Henan Prov Peoples Hosp, Peoples Hosp,Henan Eye Hosp,Henan Eye Inst, Zhengzhou, Peoples R China

来源：

ANALYTICAL BIOCHEMISTRY | 2022年 / 658卷

关键词：

CRISPR-Cas9; HRCA; DNA Detection; Visualization; NUCLEIC-ACID DETECTION; CERVICAL-CANCER; CELL-LINES; GENOME; PCR;

D O I：

10.1016/j.ab.2022.114940

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

This paper presents a novel clustered regularly interspaced short palindromic repeat (CRISPR)-associated HRCA technique (CART). During the entire detection process of CART, the target DNA is first specifically recognized and cleaved by a pair of Cas9/sgRNA complexes; then, the cleaved product is ligated into circular DNA as the template of HRCA, and the circular DNA is efficiently amplified by HRCA. Therefore, CART has the advantages of Cas9/sgRNA (single-base mismatch specificity) and HRCA (isothermal reaction temperature and high sensi-tivity). This technique has been verified by detecting various human papillomavirus (HPV) genes with numerous subtypes. In summary, this study provides a new and effective method for the detection of nucleic acids.

引用

页数：11

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