CRISPR/Cas12a and primer-assisted rolling circle amplification integrated ultra-sensitive dual-signal sensing platform for EGFR 19 detection

Liyuan Deng; Shiying Zhou; Jiangbo Dong; Yin Liu; Zhen Huang; Human Sun; Liang Jin; Danqun Huo; Changjun Hou

doi:10.1016/j.aca.2023.341755

CRISPR/Cas12a and primer-assisted rolling circle amplification integrated ultra-sensitive dual-signal sensing platform for EGFR 19 detection

Anal Chim Acta. 2023 Oct 23:1279:341755. doi: 10.1016/j.aca.2023.341755. Epub 2023 Sep 6.

Authors

Liyuan Deng¹, Shiying Zhou¹, Jiangbo Dong¹, Yin Liu¹, Zhen Huang², Human Sun¹, Liang Jin³, Danqun Huo⁴, Changjun Hou⁵

Affiliations

¹ Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, Bioengineering, College of Chongqing University, Chongqing, 400044, PR China.
² Key Laboratory of Bio-Resource and Eco-environment of Ministry of Education, College of Life Sciences, Sichuan University, SeNA Research Institute and Szostak-CDHT Large Nucleic Acids Institute, Chengdu, 610000, PR China.
³ Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, Bioengineering, College of Chongqing University, Chongqing, 400044, PR China. Electronic address: jinliang4002@126.com.
⁴ Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, Bioengineering, College of Chongqing University, Chongqing, 400044, PR China; Chongqing Key Laboratory of Bio-perception & Intelligent Information Processing, School of Microelectronics and Communication Engineering, Chongqing University, Chongqing, 400044, PR China. Electronic address: huodq@cqu.edu.cn.
⁵ Key Laboratory for Biorheological Science and Technology of Ministry of Education, State and Local Joint Engineering Laboratory for Vascular Implants, Bioengineering, College of Chongqing University, Chongqing, 400044, PR China; National Facility for Translational Medicine, Shanghai Jiao Tong University, Shanghai, 200240, PR China. Electronic address: houcj@cqu.edu.cn.

PMID: 37827629
DOI: 10.1016/j.aca.2023.341755

Abstract

Herein, we integrated CRISPR/Cas12a with primer-assisted rolling circle amplification (PARCA) to specifically detect EGFR 19 from the genome. We fused the method into fluorescent and electrochemical detection systems forming a stable and sensitive dual-signal sensing platform. The fluorescent detection system stably detected EGFR 19 in a linear range from 500 fM to 10 nM with an ultra-low background signal. The electrochemical detection system possessed a detection limit as low as 42 aM due to the introduction of nanomaterial UIO-66-NH₂. The dual-signal sensing platform showed superior performance in complex serum samples and real cell genomes and provided a flexible and dynamic approach for the ultra-sensitive detection of EGFR 19.

Keywords: CRISPR/Cas12a; Dual-signal sensor platform; EGFR 19; Rolling circle amplification (RCA).

MeSH terms

Biosensing Techniques* / methods
CRISPR-Cas Systems* / genetics
Coloring Agents
ErbB Receptors / genetics
Nucleic Acid Amplification Techniques / methods

Substances

Coloring Agents
ErbB Receptors