Alb-uPA-teton lentiviral vector and preparation method and application thereof

Abstract

The invention relates to an Alb-uPA-teton lentiviral vector and a preparation method and application thereof. The Alb-uPA-teton lentiviral vector contains an albumin promoter PAlb and a urokinase type plasminogen activator gene uPA which are reversely connected, wherein the albumin promoter PAlb is used for regulating the expression of a tetracycline trans-activator rtTA, and the urokinase type plasminogen activator gene uPA is regulated by a PTight promoter to express. The Alb-uPA-teton lentiviral vector has the advantages that the ability of establishing a system is realized, and the expression of uPA in an NOD mouse is detected.

Description

technical field [0001] The invention belongs to the field of gene function, specifically relates to an Alb-uPA-teton lentiviral vector, and also relates to a preparation method and application of the vector. Background technique [0002] In addition to humans and primates, human hepatitis virus does not have a natural small animal infection host, which seriously hinders the research on the mechanism of human hepatitis virus infection and antiviral drugs. At present, only chimpanzees are the best in vivo experimental model for studying human hepatitis virus infection, but the use of protected rare primates is subject to many restrictions such as ethics and funding. Therefore, it is an urgent problem to establish cheap and easy-to-obtain small animal models. The mouse animal model developed in recent years that can carry human liver tissue provides a good platform for the research on the mechanism of hepatitis virus infection and antiviral drugs. The study found that the uro...

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Problems solved by technology

However, the urokinase-type plasminase transgenic mice obtained by transgenic methods have high neonatal mortality and difficult screening, which greatly hinders the construction, promotion and use of human chimeric liver mouse models

[0003] The Chinese patent with the publication number CN102628063A discloses the PAlb-uPA lentiviral vector, which can overexpress the urokinase-type plasminogen activator uPA at the mouse cell level, but has been verified by a large number of animal prokaryotic injection experiments. The offspring of the transgenic mouse (founder0) failed to screen out the transgenic mouse carrying the target fragment, that is, the biological efficacy of the target gene could not be expressed in the mouse

The reasons may be as follows: (1) The constructed lentiviral plasmid carrying the target gene collapsed after in situ injection into the fertilized egg into the mouse, and could not be completely or completely re-inserted randomly into the mouse genome (2) The constructed lentiviral plasmid carrying the target gene was in situ injected into the mouse fertilized egg cells, and integrated into the mouse genome in a random manner. The design requires the expression of the biological efficacy of the target fragments; (3) the possible arrangement sequence has not changed, but the arrangement of the target fragments is affected by the spatial structure of the genome, which interferes with the expression of the target fragments

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