Issue 12, 2022
From the journal:

Analyst

Probing low abundant DNA methylation by CRISPR-Cas12a-assisted cascade exponential amplification†

Liangliang Zhang,‡ab   Xianxian Zhao,‡a   Xiaolin Hu,ab   Yi Zhang,c   Ruining Liu,a   Hai Peng,a   Youhao Chen,d   Hong Zhang*ae  and  Yang Luo ORCID logo *af  

* Corresponding authors

a Center of Smart Laboratory and Molecular Medicine, School of Medicine, Chongqing University, Chongqing 400030, P.R. China
E-mail: luoy@cqu.edu.cn, annicq@163.com

b Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, College of Bioengineering, Chongqing University, Chongqing 400044, P.R. China

c Zhejiang Provincial People's Hospital, Hangzhou, PR. China

d Department of Orthopaedics, Three Gorges Hospital, Chongqing University, Chongqing, PR. China

e Department of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, P.R. China
E-mail: annicq@163.com

f Department of Clinical Laboratory, Fuling Hospital, Chongqing University, Chongqing 408099, P.R. China
E-mail: luoy@cqu.edu.cn

Abstract

Aberrant DNA methylation plays a pivotal role in tumor development and metastasis, and is regarded as a valuable non-invasive cancer biomarker. However, the sensitive and accurate quantification of DNA methylation from clinical samples remains a challenge. Herein, we propose an easy-to-operate Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas system Assisted Methylation (CAM) approach for the sensitive detection of DNA methylation through the integration of rolling circle amplification and CRISPR-Cas12a-assisted cascade amplification. Briefly, bisulfite was employed to prepare the clinical samples so that the methylated DNA sequences trigger the subsequent triple signal amplifications, whilst the normal counterparts do not. The triple signal amplification procedure consists of methylated DNA sequence-based rolling circle amplification for a preliminary signal enhancement, a nicking enzyme-initiated target cleavage for a secondary amplification, and CRISPR-Cas12a enzyme-mediated trans-cleavage for a tertiary signal enhancement. This proposed approach reveals high sensitivity, which can even distinguish as low as 0.01% methylation levels from mixtures, paving the way towards the acceleration of methylation-based cancer diagnostics and management.

Graphical abstract: Probing low abundant DNA methylation by CRISPR-Cas12a-assisted cascade exponential amplification

About
Cited by
Related
Download options Please wait...

Supplementary files

Article information

DOI
https://doi.org/10.1039/D2AN00170E
Article type
Paper
Submitted
26 Jan 2022
Accepted
02 May 2022
First published
03 May 2022

Analyst, 2022,147, 2655-2661

Permissions

Request permissions

Probing low abundant DNA methylation by CRISPR-Cas12a-assisted cascade exponential amplification

L. Zhang, X. Zhao, X. Hu, Y. Zhang, R. Liu, H. Peng, Y. Chen, H. Zhang and Y. Luo, Analyst, 2022, 147, 2655 DOI: 10.1039/D2AN00170E

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements