Detection of small-sized DNA fragments in a glassy nanopore by utilization of CRISPR-Cas12a as a converter system. Issue 5 (10th February 2022)

• Record Type: Journal Article
• Authors: Zhang, Shumin; Liu, Minyi; Cui, Haofa; Ziaee, Muhammad Asad; Sun, Rongwei; Chen, Liting; Chen, Daqi; Garoli, Denis; Wang, Jiahai
• Is Part Of: Analyst. Volume 147:Issue 5(2022)
• Page Start: 905
• Page End: 914
• Publication Date: 2022-02-10
• Subjects: Chemistry, Analytic -- Periodicals; 543
• DOI: 10.1039/d1an02313f
• Languages: English
• ISSNs: 0003-2654

Abstract

Abstract : Small-sized DNA fragments were converted into relatively large DNA tetrahedrons based on the CRISPR-Cas12a technology. This conversion approach enlarged the signal of small targets, make it easier and more reproducible for glassy nanopores to detect. Abstract : The fabrication of nanopores with a matched pore size, and the existence of multiple interferents make the reproducible detection of small-sized molecules by means of solid-state nanopores still challenging. A useful method to solve these problems is based on the detection of large DNA nanostructures related to the existence of small-sized targets. In particular, a DNA tetrahedron with a well-defined 3D nanostructure is the ideal candidate for use as a signal transducer. Here, we demonstrate the detection of an L1-encoding gene of HPV18 as a test DNA target sequence in a reaction buffer solution, where long single-stranded DNA linking DNA tetrahedra onto the surface of the magnetic beads is cleaved by a target DNA-activated CRISPR-cas12 system. The DNA tetrahedra are subsequently released and can be detected by the current pulse in a glassy nanopore. This approach has several advantages: (1) one signal transducer can be used to detect different targets; (2) a glassy nanopore with a pore size much larger than the target DNA fragment can boost the tolerance of the contaminants and interferents which often degrade the performance of a nanopore sensor.