An electrochemical method based on CRISPR-Cas12a and enzymatic reaction for the highly sensitive detection of tumor marker MUC1 mucin

• Published in: Analyst (London) 2024-07, Vol.149 (15), p.392-3927
• Main Authors: Jin, Zhenhuan, Xiao, Wei, Shen, Lin, Shi, Xiaoxue, Li, Jianping
• Format: Article
• Language: English
• Subjects: Biomarkers; Biosensors; Cancer; CRISPR; Glassy carbon; Glucose oxidase; Hydrogen peroxide; Iron oxides; Oxidation; Tumors
• ISSN: 0003-2654; 1364-5528; 1364-5528
• DOI: 10.1039/d4an00595c

Anti-cancer therapy is crucial in cancer prevention and anti-cancer, and thus, highly sensitive methods for detecting cancer biomarkers are essential for cancer early diagnosis. Herein, an electrochemical aptamer biosensor based on the CRISPR-Cas12a system was constructed for the detection of cancer tumor biomarker MUC1 mucin. The sensitivity was significantly prompted by enzyme-catalyzed signal amplification, and the selectivity was improved by the dual recognition of the aptamer to MUC1 and crRNA-Cas12a system to the aptamer. Glucose oxidase (GOD) was loaded on the surface of magnetic Fe 3 O 4 @Au (MGNP) via probe single-stranded DNA (pDNA) with the terminal modification of mercapto (-SH) to form GOD-pDNA/MGNP. The corresponding aptamer of MUC1 (MUC1 Apt) binds to its complementary ssDNA (cDNA) to form the activator Apt/cDNA, which is specifically recognized by crRNA-Cas12a and excites the trans -cleavage function of Cas12a, thus in turn trans -cleaves pDNA and detaches GOD from the magnetic particles. The magnetic beads were separated and transferred into a glucose solution, and the oxidation current of H 2 O 2 produced by the catalytic reaction of GOD was measured on a Pt-modified magnetically-controlled glassy carbon electrode, resulting in an indirect determination of MUC1. The current change was linear with the logarithm of MUC1 concentration in the range from 1.0 × 10 −17 g mL −1 to 1.0 × 10 −10 g mL −1 . The detection limit was as low as 7.01 × 10 −18 g mL −1 . The method was applied for the detection of MUC1 in medical samples. An electrochemical aptamer biosensor based on the CRISPR-Cas12a system was constructed for the detection of cancer tumor biomarker MUC1 mucin, the sensitivity was significantly improved by employing the enzyme catalytic reaction.