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

A technology for regulating expression and mouse models, applied in the biological field, can solve the problems of limited application, low reproduction rate, and low transplantation success rate, and achieve the effects of shortening the experimental cycle, overcoming low fertility rate, and strong induction performance

Active Publication Date: 2013-01-09
FOURTH MILITARY MEDICAL UNIVERSITY
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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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  • Constructing method of mouse model for RdRp controllable express and in vivo observation
  • Constructing method of mouse model for RdRp controllable express and in vivo observation
  • Constructing method of mouse model for RdRp controllable express and in vivo observation

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Embodiment Construction

[0046] The invention provides a method for constructing a mouse model in which the expression of RdRp can be regulated and observed in vivo. First, the RdRp gene was cloned into a specific expression vector, and then the single transgenic mouse with regulated expression of RdRp was crossed with the Cre / Lap double transgenic mouse, and the RdRp / Lap gene with strong reproductive ability, low background expression and strong inducibility was screened. Cre / Lap triple transgenic mice, the selected mice can strictly regulate the expression of RdRp through dox induction, and the single transgenic mice with regulated expression of RdRp can be prepared by transgenic technology, and RdRp is not expressed when the mice are fed normally. After being induced by dox feeding, RdRp began to be expressed in the liver; the effect of RdRp inhibitors could be visually observed by mouse in vivo imaging technology; finally, the population was expanded by cross breeding of RdRp / Cre / Lap triple transge...

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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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85
Inventor 尹文孙梦宁赵亚吕欣雷迎峰杨敬康健马玉兰海云姚敏
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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