High-efficiency screening method for RNA-targeting drugs using nanopores

Patent metadata

Field	Value
Publication number	US-12560588-B2
Application number	US-202318191388-A
Country	US
Kind code	B2
Filing date	Mar 28, 2023
Priority date	Sep 28, 2020
Publication date	Feb 24, 2026
Grant date	Feb 24, 2026

Abstract

The disclosure relates to nanopore-based technology for screening of drugs against target RNA. According to the screening method of the disclosure, drugs targeting RNA can be efficiently screened even with a very small amount sample of picomolar concentration. The discovered drugs targeting RNA can be used in the treatment of diseases associated with RNAs.

First claim

What is claimed is:

1 . A method for screening an RNA-targeting drug, comprising:
detecting the presence of multi-level current signals and/or a change in dwell time of those signals associated with unzipping and translocation of a target RNA through a nanopore both before and after treating the target RNA with a candidate substance expected to bind to the target RNA; and selecting the candidate substance as a drug to be bound to the target RNA when there is a change in the presence of the multi-level current signals and/or the change in the dwell time of those signals.

2 . The method of claim 1 , wherein in the detecting step, three-dimensional structural change of the target RNA induced by binding of the candidate substance to the target RNA detected as the presence of the multi-level current signals and/or the change in the dwell time of those signals.

3 . The method of claim 2 , wherein the three-dimensional structural change of the target RNA delays the dwell time of the multi-level current signals associated with unzipping and translocation of the target RNA through the nanopore.

4 . The method of claim 1 , wherein the change in the presence of the multi-level current signals refers to an appearance of the multi-level current signals following treatment of the candidate substance, and the change in the dwell time of the multi-level current signals refers to a delay in the dwell time following the treatment of the candidate substance.

5 . The method of claim 1 , wherein the target RNA itself forms at least one stem-loop or a triplex structure.

6 . The method of claim 5 , wherein the target RNA comprises bacterial riboswitch or viral RNA promoter.

7 . The method of claim 6 , wherein the bacterial riboswitch is any one selected from the group consisting of purine riboswitch, lysine riboswitch, cyclic diGMP riboswitch, glmS riboswitch, TPP riboswitch, and FMN riboswitch.

8 . The method of claim 1 , wherein a protein that generates the nanopore is any one selected from the group consisting of a-hemolysin, ClyA, aerolysin, lysenin, CsgG, FhuA, FraC, MspA, PlyAB, Phi29, PA63, and OmpG.

9 . The method of claim 1 , wherein the RNA-targeting drug is an anticancer agent, a metabolic disease treatment, a degenerative disease treatment, a cardiovascular disease treatment, a lung disease treatment, an immune disease treatment, an antibacterial agent, an antibiotics, or an antiviral agent.

Assignees

• Korea Res Inst Bioscience & Biotechnology

Inventors

• Chi Seung-Wook
• Lee Mi-Kyung
• Lee Donghwa
• Oh Sohee

Classifications

• C12N2310/531

  Stem-loop; Hairpin · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title

• C12N2320/10

  in screening processes · CPC title

• C12N2310/16

  Aptamers · CPC title