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siRNA for inhibiting influenza virus-related gene and application

A gene and D gene technology, applied in the field of gene preparation and screening of anti-influenza drugs, can solve the problems of wrong judgment of gene function and weak gene inhibition effect, and achieve the effects of increasing drug concentration, convenient use and inhibiting virus replication.

Inactive Publication Date: 2013-07-24
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Prior to this, the complementary characteristics of antisense RNA and target mRNA sequence were used to inhibit the occurrence of its phenotype. However, due to the weak inhibitory effect of antisense RNA on endogenously expressed genes, some transitional phenotypes are often produced, which is easy to cause. Causes errors in the judgment of gene function. At present, there is only one drug Vitravene that has been approved for clinical treatment.

Method used

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  • siRNA for inhibiting influenza virus-related gene and application
  • siRNA for inhibiting influenza virus-related gene and application
  • siRNA for inhibiting influenza virus-related gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0148] Example 1 Regulates gene expression in host cells and inhibits influenza virus replication

[0149] 1. siRNA (small interfering RNA) silences gene expression in host cells and inhibits influenza virus replication

[0150] (1) siRNA molecular design

[0151] In this example, caveolin 2 gene, GeneID: 100125375; surface active protein D, GeneID: 397198 were selected, siRNA molecules were designed using Dharmacon online design software, and caveolin 2 gene-specific siRNA interference sequences were designed; and "siRNA- Caveolin" randomly scrambled sequence was used as a control. Design the siRNA interference sequence specific to the surfactant protein D gene; and use "siRNA-surfactant protein D" randomly scrambled sequence as a control. The main sequence of the designed siRNA double-strand interference molecule is 19 bases, two UUs are added to the 3' end of the sense strand, and two TTs are added to the 3' end of the antisense strand.

[0152] The designed sequence is ...

Embodiment 2

[0199] Example 2 Regulation of gene expression in host cells significantly improves the antiviral effect of influenza virus drugs

[0200] In this example, by regulating protein expression in canine kidney cells and then infecting with H3N2 swine influenza virus, by adding amantadine to the cell culture medium, the enhanced effect of amantadine on the anti-influenza virus of regulating the protein in the cells was studied. Amantadine inhibits virus replication by interfering with the ion channel of the influenza virion M2 protein.

[0201] 1 Silencing caveolin 2 in canine kidney cells significantly enhanced the inhibitory effect of amantadine on influenza virus

[0202] (1) Silencing the expression of caveolin 2 in canine kidney cells

[0203] Concrete steps are with embodiment 1.

[0204] (2) Amantadine preparation

[0205] Amantadine hydrochloride was purchased from Alfa Aisha (Tianjin) Chemical Co., Ltd., prepared with normal saline to 20 mg / ml, sterilized by filtration,...

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Abstract

The invention discloses siRNA and a recombinant vector by aiming at caveolin 2 and surfactant protein D; a construction method of a recombinant vector containing a transcription factor A and paraoxonase 3 transcription box, a corresponded cell model and a method for screening medicines. The invention has the advantages that the provided siRNA can reduce mRNA or protein expression of mammal cells after transfection by more than 60%; and the siRNA and the recombinant vector can be used for treating influenza by gene or performing nucleic acid immunization by aiming at influenza virus. The provided cell model can be used for virus research, and especially relates to swine influenza virus pathogenesis researches, and can be used for establishing a model for screening antiviral drugs, especially an anti-influenza medicine cell model.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to siRNA for inhibiting expression of caveolin 2 and surface active protein D genes, nucleotide sequence of siRNA, recombinant vector encoding the siRNA, and encoding transcription factor A and paraoxon phosphatase 3 Recombinant vectors and their respective genes in the preparation and screening of anti-influenza drugs. Background technique [0002] The process of virus invasion into the host is accompanied by changes in host gene expression, which ultimately determine the fate of infected cells and viruses. After the virus infects the host, it needs to use the metabolic process of the host cell to complete the replication of the virus itself. During this process, the virus will inevitably regulate the expression of the genome in the host cell, inhibit the expression of antiviral genes, and enhance the expression of genes that are beneficial to virus replication. Therefore, after the v...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63C12N15/79C12N5/10C12Q1/70C12Q1/48C12Q1/02A61K48/00A61P31/16
CPCC12N9/16C12N15/1131C12N15/85C12N2310/14C12Y301/08001G01N33/5026G01N33/5044G01N2500/10
Inventor 马志永史子学魏建超邵东华王少辉李蓓蓓晏文君朱紫祥
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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