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Targeted epigenome regulated and controlled hepatitis B virus (HBV)-resisting molecule HBx1C and application thereof

A hepatitis B virus, gene regulation technology, applied in the field of genes, can solve the problems of rebound, death due to pathological consequences, etc., and achieve the effect of strong infectivity

Inactive Publication Date: 2018-05-11
FUDAN UNIV
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  • Abstract
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Problems solved by technology

However, the above-mentioned drugs will have a strong rebound phenomenon after stopping the drug, causing serious pathological consequences and even death

Method used

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  • Targeted epigenome regulated and controlled hepatitis B virus (HBV)-resisting molecule HBx1C and application thereof
  • Targeted epigenome regulated and controlled hepatitis B virus (HBV)-resisting molecule HBx1C and application thereof
  • Targeted epigenome regulated and controlled hepatitis B virus (HBV)-resisting molecule HBx1C and application thereof

Examples

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Embodiment Construction

[0021] 1. Screening and construction of HBV stably transfected cell lines

[0022] The present invention uses pcDNA3.1 to construct the 1.3 ploidy of the whole HBV genome. After transfecting HepG2 cells, the HBV virus can be replicated autonomously. After G418 screening, the cell strain HepG2-HBV 1.3 that stably secretes virus particles and marker antigens is obtained, and the cell strain The HBV virus relied on is the C genotype representative of the strains circulating in East Asia.

[0023] 2. Design and synthesis of HBV X gene promoter specific binding

[0024] By using ZF Tools Ver3.0 software to predict, there are 3 CpG islands in the 3.2 kb HBV genome. Island 1 is 55-281 bp in size and contains the ATG start codon of the surface antigen; island 2 is 1228-1663 bp in size and contains enhancer 1 and X gene promoter; island 3 is 2295-2446 bp in place and contains polymerase ATG start code; CpG island 2 contains more CG sequences, and the HBVX gene promoter gene regulator...

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Abstract

The invention belongs to the technical field of genetic engineering and in particular relates to a targeted epigenome regulated and controlled hepatitis B virus (HBV)-resisting molecule HBx1C and application thereof. A brand-new protein molecule HBx1C gene, which has a targeted specific affinity interaction effect on an HBV gene and can be used for carrying out epigenetic methylation modificationon DNA (Deoxyribonucleic Acid) of the HBV gene, is designed according to bioinformatics, epigenetic inheritance engineering and an immunology principle; an adenovirus vector is introduced into cells,and a targeted and specific protein molecule aiming at an HBV can be efficiently expressed; the methylation chemical modification on the virus gene also can be generated. In vivo and vitro experimentsprove that the targeted epigenome regulated and controlled HBV-resisting molecule HBx1C has a function of efficiently inhibiting HBV surface antigen expression and a treatment effect of resisting theHBV is realized; the targeted epigenome regulated and controlled HBV-resisting molecule HBx1C can be used for preparing a medicine for treating hepatitis B in epigenetic genetic engineering and has awide application prospect.

Description

technical field [0001] The invention belongs to the field of gene technology, and in particular relates to an anti-hepatitis B virus silencing molecule targeted to epigenetic regulation and application thereof. Background technique [0002] About 300 million people in China suffer from liver diseases, mainly including viral hepatitis (hepatitis B infected with hepatitis B virus (HBV) and hepatitis C infected with HCV, and hepatitis B infected by HBV is more common in my country), non- Alcoholic fatty liver disease and alcoholic liver disease. Hepatitis, liver cirrhosis, and liver cancer have brought huge challenges to Chinese people's health and social economy. [0003] During the pathogenic process of HBV, it usually presents a typical three-step pathological process of chronic hepatitis, liver cirrhosis and liver cancer. Mutations at the level of chromosome and gene sequence can occur in the body, including chromosome deletion and rearrangement, aneuploidy, gene expansion ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/861A61K48/00A61P31/20
CPCA61K48/005C12N9/1007C12N15/86C12N2710/10043
Inventor 朱乃硕李雪
Owner FUDAN UNIV
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