shRNA capable of inhibiting replication and infection of Japanese encephalitis virus and application thereof

A technology of Japanese encephalitis virus and template sequence, applied in the field of genetic engineering

Inactive Publication Date: 2011-04-06
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 capable of inhibiting replication and infection of Japanese encephalitis virus and application thereof
  • shRNA capable of inhibiting replication and infection of Japanese encephalitis virus and application thereof
  • shRNA capable of inhibiting replication and infection of Japanese encephalitis virus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation of embodiment 1 shRNA

[0035] (1) Design and synthesis of oligonucleotides

[0036] Taking JEV's E, NS3, NS4b gene mRNA sequence as the target sequence ( figure 1 ), using Ambion’s siRNA design software, through homology analysis, designed and synthesized 4 for 1553~1571bp (E1), 1679~1697bp (E2), 6083~6101bp (NS3) and 7350~7368bp (NS4b) of the JEV gene, respectively. 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 templa...

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Abstract

The invention belongs to the technical field of genetic engineering and discloses shRNA capable of inhibiting replication and infection of Japanese encephalitis virus and application thereof. The shRNA takes a highly-conserved sequence in a Japanese encephalitis virus genome as a target sequence which is SEQIDNO.9, SEQIDNO.10, SEQIDNO.11 or SEQIDNO.12. The shRNA eukaryotic expression plasmid can be obtained by connecting a template of the shRNA with a linearized vector pGPU6 / GFP / Neo and respectively aims at Japanese encephalitis virus E and NS3 or NS4b gene mRNA sequence. The shRNA and the shRNA eukaryotic expression plasmid provided by the invention can be applied to a medicament for treating and / or preventing JEV (Japanese Encephalitis Virus) infection and provides a novel selectable medicament for treating and preventing the JEV infection.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to shRNA capable of inhibiting the replication and infection of Japanese encephalitis virus and its application. Background technique [0002] 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 has also been continuously enhanced. [0003] JE virus belongs to the genus Flaviviridae of the family Flaviviridae. Virus particles are icosahedron symmetrical, with a dense lipoprotein envelope surrounding the nucleic acid. The JEV genome is a single-stranded positive-sense RNA molecule, which is i...

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
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