A novel nanoparticle surface-constrained CRISPR-Cas12a 3D DNA walker-like nanomachines for sensitive and stable miRNAs detection

• Published in: Analytica chimica acta 2023-04, Vol.1251, p.340950-340950, Article 340950
• Main Authors: Wang, Xin, Mu, Xiaomei, Li, Jinshen, Liu, Guang, Zhao, Shulin, Tian, Jianniao
• Format: Article
• Language: English
• Subjects: 3D DNA walker-like; AuNPs; Biosensing Techniques; CRISPR-Cas Systems; CRISPR-Cas12a; DNA - genetics; Gold; Metal Nanoparticles; MicroRNAs - genetics; miRNA-21; Nucleic Acids; RCA; SNA
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2023.340950

The CRISPR-Cas system has broad prospects as a new type of nucleic acid signal amplification technology based on the trans-cleavage activity of Cas12a to single-stranded DNA, but the trans-cleavage reaction efficiency is relatively low in solution. In order to overcome this negative factor, a new 3D DNA nanomachine whose CRISPR-Cas12a is limited to the surface of nanoparticles is used for sensitive and stable detection of miRNA. By loading Cas12a activator onto spherical nucleic acid (SNA), the CRISPR-Cas12a activator system on the surface of Au nanoparticles (AuNPs) acts as a walker to carry out continuous recognition-walking-cutting reaction on the surface of AuNPs, which enhances the trans-cleavage activity of Cas12a to SNAs. Benefiting from the confinement effect of spherical nucleic acids surface, a 3D DNA nanomachine has been developed for the detection of miRNA-21, which has achieved high sensitivity and accuracy, and the detection limit is able to reach 8.0 pM. This new 3D DNA walker-like nanomachine provided another insight for future bioanalysis and early clinical diagnoses of disease and liquid biopsy. A 3D DNA nanomachine with CRISPR-Cas12a confined to the surface of nanoparticles was developed and used for sensitive and stable detection of miRNAs. [Display omitted] •A novel 3D DNA walker-like structure based on CRISPR/Cas system and spherical nucleic acids (SNA).•The CRISPR/Cas as the walking leg on spherical nucleic acids.•The walking cleavage efficiency of Cas12a and accelerates shear recognition response because of the confinement effect of spherical nucleic acids surface. Because of the confinement effect the walking cleavage efficiency is accelerated.