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. This journal is

Detection of small-sized DNA fragments in a glassy nanopore by utilization of CRISPR-Cas12a as a converter system / Zhang, Sm; Liu, My; Cui, Hf; Ziaee, Ma; Sun, Rw; Chen, Lt; Chen, Dq; Garoli, D; Wang, Jh. - In: ANALYST. - ISSN 0003-2654. - 147:5(2022), pp. 905-914. [10.1039/d1an02313f]

Detection of small-sized DNA fragments in a glassy nanopore by utilization of CRISPR-Cas12a as a converter system

Garoli D;
2022

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. This journal is
2022
147
5
905
914
Detection of small-sized DNA fragments in a glassy nanopore by utilization of CRISPR-Cas12a as a converter system / Zhang, Sm; Liu, My; Cui, Hf; Ziaee, Ma; Sun, Rw; Chen, Lt; Chen, Dq; Garoli, D; Wang, Jh. - In: ANALYST. - ISSN 0003-2654. - 147:5(2022), pp. 905-914. [10.1039/d1an02313f]
Zhang, Sm; Liu, My; Cui, Hf; Ziaee, Ma; Sun, Rw; Chen, Lt; Chen, Dq; Garoli, D; Wang, Jh
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/1315973
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