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FAT10 gene siRNA recombination analogue virus as well as preparation method and application thereof

A technology of FAT10 and gene, applied in the field of simulated virus, can solve the problems of unstable effect, low plasmid transfection efficiency, complicated preparation process, etc., and achieve the effect of simple and time-saving preparation method, good development and application prospects, and avoiding damage to irrelevant cells

Inactive Publication Date: 2012-07-25
ARMY MEDICAL UNIV
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  • Abstract
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Problems solved by technology

Although RNAi mediated by viral vectors has attracted much attention in recent years, the use of viral vectors can solve the problems of low plasmid transfection efficiency, unstable effect, and certain types of cells cannot be transfected, but it also has problems such as immunogenicity and virality. Safety issues such as recombination, as well as defects such as complicated preparation process, limited capacity, and low titer of virus produced

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  • FAT10 gene siRNA recombination analogue virus as well as preparation method and application thereof
  • FAT10 gene siRNA recombination analogue virus as well as preparation method and application thereof
  • FAT10 gene siRNA recombination analogue virus as well as preparation method and application thereof

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Embodiment Construction

[0031] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. The experimental method that does not indicate specific conditions in the preferred embodiment is usually according to conventional conditions, such as described in the Molecular Cloning Experiment Guide (Third Edition, J. Sambrook et al., translated by Huang Peitang, etc., Science Press, 2002) conditions, or as recommended by the manufacturer.

[0032] 1. Preparation and identification of FAT10 gene siRNA recombinant simulated virus

[0033] 1. Construction of FAT10 gene siRNA recombinant expression vector

[0034] (1) Synthesis of shRNA transcription template DNA

[0035] According to the literature (Liu Tiande et al. Study on the inhibitory effect of siRNA on the abnormal expression of FAT10 gene in liver cancer cells. New Medicine, 2009, 40(6): 358) reported FAT10 gene siRNA sequence: sense strand: 5′-gcuc-aguggcacaagugaatt-3′ (SEQ ...

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Abstract

The invention relates to an analogue virus, in particular to an FAT10 gene siRNA recombination analogue virus as well as a preparation method and application thereof. The analogue virus is a compound of a fusion peptide and an FAT10 gene siRNA recombination expression vector, wherein the fusion peptide is formed by connecting an RGD peptide and a cell-penetrating peptide HIV-Tat49-57; and the FAT10 gene siRNA recombination expression vector controls the expression of the FAT10 gene siRNA through an alpha fetoprotein promoter. The analogue virus can target hepatocarcinoma cells and limit the expression of the FAT10 gene siRNA in the hepatocarcinoma cells and cause silent expression of the FAT10 gene in the hepatocarcinoma cells because of the specificity to avoid the damage to the irrelevant cells, and has the advantages of high transfection efficiency, simple and time-saving preparation method, and the like. The analogue virus can be used for preparing anti-cancer drugs and has good prospect in development and application in the field of the gene therapy of liver cancers.

Description

technical field [0001] The invention relates to a simulated virus, in particular to a FAT10 gene siRNA recombinant simulated virus, and also relates to a preparation method and application of the simulated virus. Background technique [0002] Liver cancer is a common malignant tumor, ranking fifth in the incidence of malignant tumors, and its exact pathogenesis has not yet been fully understood. How to block the malignant transformation of chronic liver disease to liver cancer, or intervene in the early stage of liver cancer, has become the key to further improve the treatment effect of liver cancer. [0003] FAT10, also known as diubiquitin, belongs to the ubiquitin-like modified proteins (UBLs) in the ubiquitin protein family. It is a protein with a molecular weight of 18 kDa and 165 amino acid residues. domains merged. FAT10 plays a very important role in the regulation of the cell cycle, and it has been proven that it can non-covalently bind to the human spindle aggreg...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/63C12N15/113A61K48/00A61P1/16A61P35/00
Inventor 杨曌吴玉章
Owner ARMY MEDICAL UNIV
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