Constructing method of mouse model for RdRp controllable express and in vivo observation

Abstract

The invention discloses a construction method a mouse model for RdRp controllable express and in vivo observation, which comprises the following steps: carrying out hybridizing by respectively taking a transgenic mouse of an RdRp controllable express vector, which is recombined by a genome and a Cre/Lap double transgenic mice as parents; selecting filial generation of the parent with strong fertility; authenticating individuals having Cre, Lap and RdRp genes contained in the genome by using PCR (Polymerase Chain Reaction) amplification and taking the individuals as a filial generation of an F1 generation of a Cre/Lap/RdRp three-transgenic mice; hybridizing the filial generation of the F1 generation with a parent of a normal mouse; and screening filial generation individuals which have strong fertility and have the Cre, Lap and RdRp genes contained in the genome to be continuously mated with the normal mouse, breeding and expanding into groups until obtaining a Cre/Lap/RdRp stably-genetically model mouse.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the construction of a novel HCV infection animal model, in particular to a construction method and application of a mouse model in which RdRp can regulate expression and observation. Background technique [0002] Hepatitis C virus (HCV) is the main pathogen causing chronic liver disease. There are about 170-200 million infected people in the world, and about 50 million HCV infected people in my country. After being infected by HCV, 50% to 85% of infected persons develop chronic hepatitis, 20% to 30% of infected persons develop liver cirrhosis, and some develop liver cancer. HCV seriously endangers human health and has become a global and important public health problem. So far, there is still a lack of effective anti-HCV treatment measures, and no vaccine is available. [0003] The full length of the HCV genome is about 9.6kb, containing a single open reading frame (ORF), which encod...

AI Technical Summary

Problems solved by technology

Chimpanzees are so far the only animal model suitable for HCV infection and supporting its replication. However, chimpanzees are not produced in my country and must be imported and strictly restricted by the "International Convention". Moreover, the reproduction rate is low, the number is limited, and the cost is expensive. Therefore, its source applications are extremely limited

Although studies have shown that tree shrews can form symptoms of acute HCV infection, the virus replication and expression levels after HCV infection in tree shrews are not high, and it is difficult to establish a model of HCV chronic infection. limit their application

[0007] The disadvantages of the human-mouse liver chimeric mouse model are limited sources of human hepatocyte donors, difficult transplantation techniques, low success rate of transplantation, and not suitable for large sample screening and statistical analysis

[0008] The main problem of transgenic mice as an animal model of HCV infection is that the transgenic mice express the target gene as soon as they are passed on, which makes the body mistakenly recognize it as its own protein and make the mice in a state of immune tolerance for life, which is in a state of natural infection with HCV. The differences are large, and the resulting data and results lack credibility, which limits the evaluation of pathogenic mechanisms and antiviral drugs

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