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Application of p97 gene for therapy of virus infection and related medicine thereof

A technology for virus infection and drug treatment, which can be used in antiviral agents, medical preparations containing active ingredients, pharmaceutical formulas, etc. The effect of anti-viral infection ability

Active Publication Date: 2017-01-11
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] But the role of p97 in virus infection immunity has not been seen in the prior art

Method used

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  • Application of p97 gene for therapy of virus infection and related medicine thereof
  • Application of p97 gene for therapy of virus infection and related medicine thereof
  • Application of p97 gene for therapy of virus infection and related medicine thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0102] Example 1 p97 Negatively Regulates Type I Interferon Signaling Pathway Depends on Its ATPase Activity

[0103] 1. Experimental purpose: To determine whether p97 is involved in the signaling pathway of type I interferon response and antiviral immunity.

[0104] 2. Experimental method:

[0105] (1) Cell infection experiment: Human HEK293, HEK293T, and mouse MEF cells were purchased from the Cell Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. The cell culture method was as described in Basic Experimental Method 10. Sendai virus Sendai virus (SeV) and GFP-Vesicular stomatitis virus vesicular stomatitis virus with green fluorescence (GFP-VSV) were donated by Professor Shu Hongbing of Wuhan University. SeV and VSV-GFP are commercially available, and the transformation method of VSV-GFP is the same Construction and expression of high titer pseudotyped GFP-retroviruses by cotransfection of VSV-G geneJournal of Shanghai Jiaotong University(Me...

Embodiment 2

[0126] Example 2 Npl4 cooperates with p97 to regulate antiviral immune response

[0127] 1. Experimental purpose: p97 binds to different cofactors to participate in the regulation of different signaling pathways. The purpose of this example is to determine the cofactors bound by p97 in the regulation of antiviral immune response.

[0128] 2. Experimental method:

[0129] (1) siRNA to knock out p97, Npl4, Ufd1, p47, and FAF1 in cells: the above siRNA was synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd., the p97 siRNA sequence is shown in Table 6, and Npl4, Ufd1, p47, and FAF1 are shown in Table 7 .

[0130] Table 7 siRNA sequence

[0131]

[0132] (2) The detection of the activity of the reporter gene and the transcription level of the gene is the same as in Example 1.

[0133] (3) Construction of Npl4 vector overexpressed in cells

[0134] Extract the total cellular RNA with the method and system in Example 1, and reverse transcribe it into cDNA, and us...

Embodiment 3

[0151] Example 3 Structural analysis of p97-Npl4 complex

[0152] 1. The purpose of the experiment: through the method of structural biology, the p97 and Npl4 proteins obtained in vitro were used to assemble the complex, analyze its structure, and study the protein interaction mode.

[0153] 2. Experimental method:

[0154] (1) Expression vector construction for complex structure analysis:

[0155] HT-represents the pET28a-HisTEV vector, which is a vector constructed by the laboratory and transformed from the commercial vector pET28a (Merck). For plasmid information, see (Shi, Z., et al. Structure of the MST4in complex with MO25 provides insights into its activation mechanism. Structure 21, 449-461, 2013) Drosophila HT-Ter94-N(20-186), Drosophila HT- Npl4-UBD(1-77) uses the full-length Drosophila Ter94 and reverse-transcribed cDNA as a template, and Takara high-fidelity DNA polymerase PCR amplification was carried out with the corresponding primers under the action of the c...

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Abstract

The invention relates to the field of biotechnology and particularly relates to an application of a p97 gene for therapy of virus infection and a related medicine thereof. For the first time, the invention discloses that a p97-Npl4 compound formed by combining the p97 with an assistant factor Npl4 can promote degradation of RIG-I by promoting K48 chain ubiquitination of the RIG-I, which is mediated by RNF125, thereby inhibiting RIG-I-mediated IFN[beta] signal channel, and finally achieving the effect of reducing anti-virus infection capability. On the basis of the mentioned mechanism, an inhibitor, which can effectively block the interaction of the p97 and the Npl4, is screened out, wherein the inhibitor can promote the RIG-I-mediated IFN[beta] signal channel, thus improving the anti-virus infection capability and further achieving the effects of therapy on the virus infection. The application has great clinical and market value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of p97 gene in treating virus infection and related medicines. Background technique [0002] Viral infection can cause a strong antiviral immune response in the body. Natural immunity is the first barrier for the body to resist viral infection. It passes through pathogen recognition receptors in the body, including toll-like receptors (TLR), RIG-I RLRs) can recognize viral nucleic acids. The activated receptor can start the signal transduction pathway of downstream transcription factor NF-κB and interferon regulatory factor IRF, and produce a series of pro-inflammatory cytokines and type I interferons (type I interferons, type I IFN). The release of type I interferon can promote the transcription of downstream genes and effectively inhibit the further replication of the virus. At the same time, the secretion of type I interferon can also activate cells involved in the innat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02A61K45/00A61K31/704A61P31/12A61P31/14
Inventor 周兆才焦石
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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