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shRNA for inhibiting Japanese encephalitis virus replication and infection and application thereof

A Japanese encephalitis virus and expression plasmid technology, applied in the field of shRNA, to achieve the effect of treating JEV infection, reducing morbidity and mortality, and preventing JEV infection

Inactive Publication Date: 2012-03-21
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective treatment for JE, so prevention of JE is particularly important
However, traditional vaccine production cannot meet the growing needs of human beings. Therefore, researchers are constantly looking for new treatments

Method used

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  • shRNA for inhibiting Japanese encephalitis virus replication and infection and application thereof
  • shRNA for inhibiting Japanese encephalitis virus replication and infection and application thereof
  • shRNA for inhibiting Japanese encephalitis virus replication and infection and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The preparation of embodiment 1 shRNA

[0030] (1) Design and synthesis of oligonucleotides

[0031] Taking JEV's E, NS3, NS4b gene mRNA sequence as the target sequence ( figure 1), using Ambion’s siRNA design software, through homology analysis, respectively designed and synthesized 4 for 1553-1571bp (E1), 1679-1697bp (E2), 6083-6101bp (NS3) and 7350-7368bp (NS4b) of the JEV gene for shRNA oligonucleotide strands. TTCAAGAGA is selected for its loop structure to avoid the formation of termination signal, and the transcription termination sequence of shRNA adopts T6 structure. CACC is added to the 5' end of the sense strand template, which is complementary to the sticky end formed after digestion with BbsI; GATC is added to the 5' end of the antisense strand template, which is complementary to the sticky end formed after digestion with BamHI; if the target sequence of shRNA If the first base is not G, a G is added after the CACC of the template strand. The template ...

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Abstract

The invention belongs to the technical field of genetic engineering, and discloses shRNA for inhibiting Japanese encephalitis virus replication and infection and an application thereof. The shRNA adopts a highly conserved sequence in Japanese encephalitis virus genome as a target sequence, and the target sequence is SEQ ID NO. 10. The shRNA eukaryotic expression plasmid is obtained by connecting the shRNA template to a linearized vector pGPU6 / GFP / Neo, and is aimed at a Japanese encephalitis virus E gene mRNA sequence. The shRNA eukaryotic expression plasmid provided by the invention is applicable to the preparation of medicaments for treating and / or preventing JEV infection, and provides a new optional medicament for the treatment and prevention of JEV infection.

Description

[0001] Case Description [0002] This case is a divisional application of a Chinese invention patent application with the application number 201010507298.7 and the application date on October 14, 2010, and the name is a shRNA capable of inhibiting the replication and infection of Japanese encephalitis virus and its application. technical field [0003] The invention belongs to the technical field of genetic engineering, and relates to shRNA capable of inhibiting JEV replication and infection and application thereof. Background technique [0004] Japanese encephalitis (JE) is a serious zoonotic arboviral disease caused by Japanese encephalitis virus (JEV). The disease is very harmful to humans and is one of the most common insect-borne diseases of the human central nervous system. It is widely distributed in Asia, especially in some countries and regions in the Far East. In recent years, its popular distribution range has been continuously expanded, and its popular frequency...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/85A61K48/00A61P31/14
Inventor 沈婷陈溥言刘珂曹瑞兵周斌
Owner NANJING AGRICULTURAL UNIVERSITY