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Modified nucleic acid inhibiting micro rna, and use thereof

A nucleic acid and nucleotide technology, which is applied in the field of modified nucleic acid to inhibit microRNA, can solve the problems of adverse effects, inability to distinguish normative targets from non-normative targets, etc., and achieve the effect of preventing competitive inhibition and effectively inhibiting miRNA

Pending Publication Date: 2021-03-16
KOREA UNIV RES & BUSINESS FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this type can repress specific miRNAs but cannot distinguish between canonical targets recognized by miRNAs and non-canonical targets for repression
Therefore, when it is used as a drug, it may have adverse effects, also inhibiting all biologically desired miRNA functions

Method used

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  • Modified nucleic acid inhibiting micro rna, and use thereof
  • Modified nucleic acid inhibiting micro rna, and use thereof
  • Modified nucleic acid inhibiting micro rna, and use thereof

Examples

Experimental program
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Embodiment approach

[0059] There is no limitation on the type of miRNA according to the present invention, and according to an exemplary embodiment of the present invention, the miRNA may be miRNA-1.

[0060] In the present invention, the miRNA target site sequence may include at least 6 nucleotides, preferably 21 or less nucleotides, most preferably 6 to 8 nucleotides, but the present invention is not limited thereto.

[0061] Since the miRNA actually only recognizes at least 6 base pairs in positions 1 to 8 from its 5' end, even a sequence complementary only to the base sequence in positions 1 to 8 can sufficiently bind to the miRNA, and When the miRNA target site sequence consists of 6 to 8 nucleotides, only the canonical target recognition of the miRNA can be competitively inhibited.

[0062] In the present invention, the miRNA-inhibiting modified nucleic acid may contain at least two miRNA target site sequences consecutively, and is better because it contains two or more miRNA target site se...

Embodiment 1

[0095] Example 1. Demonstration of the effect of inhibiting the canonical seed-binding function of miRNAs by tandem target miRNA inhibitors

[0096] 1-1. Method for preparing tandem target miRNA inhibitors

[0097] Such as figure 1 As shown, 5'-UAAGGCACG-3' is a base sequence from 1 to 8 positions from the 5' end of the base sequence of the miRNA that binds to the Argonaute protein, and is the seed base of the corresponding miRNA (for example, miR-124) base sequence, and the miRNA uses this sequence to bind complementary to the target mRNA. Here, when a tandem target miRNA inhibitor (a modified nucleic acid that inhibits miRNA) is prepared by introducing two or more short 8-nt target sequences capable of complementary pairing with the miRNA seed, the miRNA seed (5'-UAAGGCACG- 3') and the 5'-UGCCUU-3' sequence of the tandem target miRNA inhibitor are fully complementary, thereby relatively inhibiting the base pairing with the actual target mRNA and inhibiting the canonical ...

Embodiment 2

[0112] Example 2. Discovery and validation of non-canonical target sites that allow wobble base pairing in miRNA seed regions

[0113] 2-1. Ago HITS-CLIP method to detect non-canonical G:A wobble pairing sites of miRNAs expressed in the cerebral cortex

[0114] As a method for analyzing miRNA targets at the transcriptome level, an experimental method called AgoHITS-CLIP has been developed and widely used. AgoHITS CLIP analysis is a method in which RNA in cells is induced by irradiating cells or tissue samples with ultraviolet light. Covalently bonded to Argonaute proteins, the resulting RNA-Argonaute complexes were isolated by immunoprecipitation using antibodies specific for Argonaute, and the isolated RNA was analyzed by next-generation sequencing. The base sequence data thus obtained can not only identify target mRNAs of miRNAs through bioinformatics analysis, but also precisely analyze their binding sites and sequences. AGO HITS-CLIP was first applied to mouse cerebral ...

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Abstract

The present invention relates to a modified nucleic acid that inhibits micro RNA, and a use thereof. The modified nucleic acid that inhibits micro RNA, according to the present invention, can stronglybond with micro RNA by consecutively arranging at least two micro RNA target sites comprising a complementarily-bonding nucleotide sequence of a minimum of six on a micro RNA seed region, and, as a result of the insertion of an abasic spacer between the micro RNA target sites, not only effectively inhibits the micro RNA, but also has the benefit of the sequence inserted in the modified nucleic acid preventing inadvertent bonding with another micro RNA, and not only canonical target but also non-canonical target recognition of micro RNA such as a nucleation bulge site and G:A or G:U wobble arrangement sites can be used, and it is possible to selectively inhibit biological functions taking place due to non-canonical target recognition of micro RNA.

Description

technical field [0001] The present invention relates to a modified nucleic acid for inhibiting microRNA and its application, more specifically, to a modified nucleic acid for inhibiting microRNA that complementarily binds to microRNA and its application. [0002] This application claims the priority and benefits of Korean Patent Application No. 10-2018-0063047 filed on May 31, 2018 and Korean Patent Application No. 10-2019-0064767 filed on May 31, 2019, and its specification and drawings The disclosure of is incorporated herein by reference in its entirety. Background technique [0003] RNA interference is a phenomenon that suppresses gene expression at the post-transcriptional level, and is widely used as a technique to suppress a desired target gene by introducing an RNA substance that causes this phenomenon into cells to artificially induce gene silencing. RNA interference is actually mediated by small RNAs consisting of about 21 nucleotides (called microRNAs (miRNAs), a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N5/077
CPCC12N15/113C12N15/63C12N15/67C12N2310/113A61K31/7088C12N2310/332C12N2310/3519C12N2310/11C12N2310/51C12N5/0657C12N2510/00
Inventor 池晟旭张恩淑石熙暎
Owner KOREA UNIV RES & BUSINESS FOUND
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