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A polyoligonucleotide molecule and its application in multi-target interference

A technology of polynucleotides and oligonucleotides, which is applied in the field of polyoligonucleotide molecules and their application in multi-target interference, can solve the problem that a single molecule cannot be guaranteed to inhibit target gene expression, endonuclease and exonuclease Highly sensitive enzymes, cumbersome synthesis steps and other issues, to achieve the effect of multi-target interference, RNAi efficiency improvement, and broad application prospects

Active Publication Date: 2022-04-15
GUANGZHOU RIBOBIO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, traditional RNAi reagents have the following deficiencies: 1) cumbersome synthesis steps and relatively high production costs; 2) highly sensitive to endonucleases and exonucleases, and low stability; 3) inhibition efficiency is not high enough to prevent Ensure that a single molecule must be able to inhibit the expression of the target gene; 4) Side effects caused by non-specific activity, the non-specific effect is mainly derived from the sense strand; 5) Difficult to introduce into cells, especially animals

Method used

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  • A polyoligonucleotide molecule and its application in multi-target interference
  • A polyoligonucleotide molecule and its application in multi-target interference
  • A polyoligonucleotide molecule and its application in multi-target interference

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Effect test

Embodiment 1

[0130] Embodiment 1, the design and synthesis of polyoligonucleotide molecule

[0131] 1. Design of Polyoligonucleotide Molecules

[0132] The polyoligonucleotide molecule of the present invention comprises at least 3 oligonucleotide sequences, the size of each oligonucleotide sequence in the polyoligonucleotide molecule is 15-50nt, and each oligonucleotide sequence starts at the 5' end The 3' end is sequentially composed of a targeting terminal fragment TTS with a size of 5-24nt, a targeted internal fragment TIS with a size of 1-20nt, and an adapter terminal fragment ATS with a size of 5-24nt. Each oligonucleotide sequence in the polyoligonucleotide molecule forms a double-stranded region with the other two sequences in the polyoligonucleotide molecule except itself through sequence complementarity, that is, the targeting terminal of each oligonucleotide The fragment TTS is complementary to the adapter terminal fragment ATS of its adjacent oligonucleotide; and the targeting ...

Embodiment 2

[0143] Example 2. Application of Polyoligonucleotide Molecules in Inhibiting Target Gene Expression in Tumor Cells

[0144] (1) Inhibition test for the same target gene

[0145] 1. Molecular inhibition test of polyoligonucleotides with different numbers of sequences

[0146] 1. Molecular inhibition test of polyoligonucleotide for different sequence numbers of p65 gene

[0147] (1) Preparation of polyoligonucleotide molecules

[0148] Three kinds of polyoligonucleotides N(R m1 +R m2 +R m3 ), namely 4(R12+R6+R12), 5(R12+R6+R12), 6(R12+R6+R12). Among them, N represents the number of sequences, m1 represents the nucleotide number of the TTS fragment of each sequence, m2 represents the nucleotide number of the TIS fragment of each sequence, and m3 represents the nucleotide number of the ATS fragment of each sequence , wherein each oligonucleotide sequence is RNA, the nucleotide sequence is shown in Table 1, and the target is mRNA. Among them, the sequence of lowercase letter...

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Abstract

The invention discloses a polyoligonucleotide molecule and its application in multi-target interference. The present invention combines CON technology with nanotechnology, and realizes multi-target interference by constructing an accurate and self-assembling nucleic acid structure, which can be used to inhibit the expression of multiple genes in the signal pathway of disease occurrence or development, or simultaneously inhibit The expression of multiple disease target genes has broad application prospects in biology, chemistry and other disciplines. It is proved by experiments that the polyoligonucleotide molecule of the present invention has the following advantages: 1) RNAi efficiency is improved; 2) chemical stability is good, nuclease degradation resistance is enhanced, and half-life is prolonged; 3) off-target rate is reduced; 4) it can form Plastic nanoparticles, the ability to import into cells is enhanced.

Description

technical field [0001] The invention belongs to the field of nucleic acid nanotechnology, and specifically relates to a polyoligonucleotide molecule and its application in multi-target interference. The molecule can efficiently regulate the expression of one or more target genes. Background technique [0002] CON (complementary oligonucleotide) technology enables artificially synthesized oligonucleotide molecules to bind to target molecules through sequence complementarity and change the biological characteristics of target molecules. CON technology includes two categories of RNAi (RNA interference) technology and ASO (antisense oligonucleotide) technology, which have been widely used in functional genomics research and are expected to become the third major treatment in addition to small molecule compounds and biological agents means. For example, mipomersen, which is used to treat homozygous familial hypercholesterolemia (FoFH), is a synthetic phosphorothioate oligonucleo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/10A61K48/00A61P35/00
CPCA61K48/005C12N15/113C12N2310/14
Inventor 张必良杨秀群丹米其·萨玛斯基
Owner GUANGZHOU RIBOBIO
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