Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

AAV-HBV recombinant virus based on S gene cleavage, and establishment method and application of hepatitis B virus mouse model

An AAV-HBV, AAV-HBV1.04 technology, applied in the direction of single-stranded DNA virus, retro DNA virus, virus, etc., can solve the problem of not being HBV cccDNA, unable to judge the role of cccDNA, etc.

Active Publication Date: 2021-04-30
WUHAN UNIV
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

All of the above models have deficiencies, either it is impossible to judge whether the cccDNA is functioning, or the formed cccDNA is a recombinant cccDNA, not the real HBV cccDNA

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • AAV-HBV recombinant virus based on S gene cleavage, and establishment method and application of hepatitis B virus mouse model
  • AAV-HBV recombinant virus based on S gene cleavage, and establishment method and application of hepatitis B virus mouse model
  • AAV-HBV recombinant virus based on S gene cleavage, and establishment method and application of hepatitis B virus mouse model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The pAAV-HBV1.2 and pAAV-HBV1.04 plasmids were constructed. The pAAV-HBV 1.04 plasmid was constructed by homologous recombination technology, see the schematic diagram figure 1 , the pAAV-HBV1.04 vector plasmid sequence is shown in SEQ ID NO.3. Select the small S region of the HBV genome rcDNA as the starting point and extend along the HBV genome rcDNA back to the starting point and then about 127 bp (homologous arm) as the end point. The obtained HBV genome DNA length divided by 3182 is 1.04 copies. The selection of the starting point and the end point can change back and forth in the small S area.

[0036] 1. Amplification of AAV linearized vector backbone fragment and HBV target fragment by PCR technology

[0037]The template used to amplify the AAV vector backbone is pAAV-TBG>hSLC10A1[NM_003049.3]:WPRE (the AAV vector backbone sequence is a commonly used sequence). The template used to amplify the HBV target fragment is the plasmid containing HBV1.3 ploid or dipl...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention belongs to the technical field of biology, and particularly relates to an AAV-HBV recombinant virus based on S gene cleavage, and an establishment method and application of a hepatitis B virus mouse model. According to the invention, pAAV-HBV1.04 plasmids are constructed by adopting a homologous recombination method, and the recombinant virus is produced by in vitro packaging through a three-plasmid co-transfection 293T cell method. The recombinant virus is used for forming an HBV chronic infection mouse model in which HBV cccDNA continuously exists through caudal vein injection of a C57BL / 6 mouse with a normal immune system. The mouse model provided by the invention can generate real HBV cccDNA and can be used as a unique transcription template to play a role. The novel HBV chronic infection mouse model constructed by the invention can be used for researching a mechanism for forming and maintaining HBV cccDNA under a long-term chronic infection condition, researching problems related to an immune system, and searching and evaluating a new anti-HBV drug.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for establishing a mouse model of AAV-HBV recombinant virus and hepatitis B virus based on S gene disruption and its application. Background technique [0002] Hepatitis B virus (HBV) is a hepatotropic DNA virus, and its genome is a partially double-stranded relaxed circular DNA (Relaxed circular DNA, rcDNA), with a total length of about 3.2kb. It is the smallest DNA virus known so far (Ganem, D. and A.M. Prince, Hepatitis B virus infection--natural history and clinical consequences. N Engl J Med, 2004.350(11):p.1118-29.). After HBV infects host cells, its rcDNA will be repaired into covalently closed circular DNA (cccDNA) by the DNA damage repair system in the host cell nucleus, and cccDNA is the template for viral transcription. Although the vaccine is effective in preventing HBV infection, it does not help about 250 million chronic hepatitis B patients worldw...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N15/36C12N15/864A01K67/027
CPCC12N7/00C12N15/86C07K14/005A01K67/0278C12N2730/10122C12N2750/14121C12N2750/14143A01K2207/15A01K2217/052A01K2227/105A01K2267/0337
Inventor 夏宇尘李聪钟有权
Owner WUHAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com