Construction method for hepatitis B virus (HBV) non-human animal model and application of HBV non-human animal model

Abstract

The invention relates to a construction method for a hepatitis B virus (HBV) non-human animal model and application of the HBV non-human animal model. The construction method comprises the step: constructing a transgenic animal into which a first fusion gene segment and a second fusion gene segment are transferred, wherein the first fusion gene segment includes an inducible response element, a promoter, a nuclear localization signal and site-specific recombinase; the inducible response element can make the site-specific recombinase expressed under the condition that an inducer is present; thesecond fusion gene fragment includes an HBV gene and a reporter gene; the reporter gene is divided into two segments, and the two segments are fused with an N terminal and C terminal of the HBV gene respectively; two ends of the second fusion gene fragment are each further provided with a recognition site for the site-specific recombinase; under the action of the site-specific recombinase, the second fusion gene fragment can be cyclized, and the two segments of the reporter gene are fused into a complete active segment; and when the transgenic animal is injected with the inducer, HBV cccDNA can be generated.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing an HBV non-human animal model and its application. Background technique [0002] Hepatitis B virus (HBV) is a unique double-stranded DNA virus with a reverse transcription process, belonging to the Hepadnaviridae family. The total length of its genome is about 3.2kb, encoding seven functional proteins: HBV core antigen (HBcAg), e antigen (HBeAg), surface antigen (HBsAg), polymerase (pol) and X protein (HBx). During the process of infecting host cells, the virus will release viral nucleic acid (RC-DNA) in the cytoplasm, and further form covalently closed circular DNA (cccDNA) in the nucleus or directly integrate into the host genome. Among them, cccDNA exists in the form of minichromosomes, directs the synthesis of pre-genomic RNA (pre-genomic RNA), and controls the life cycle of viruses through a series of epigenetic regulation (Chen et al., 2015; Seeger an...

AI Technical Summary

Problems solved by technology

Not only that, but the above-mentioned various mouse models have a biggest defect: there is no formation of cccDNA in mouse liver cells, so the model can only study the interaction between HBV viral proteins and host factors, and cannot reflect the true biological characteristics of cccDNA

However, the Cre-Loxp-mediated cccDNA mouse model will be quickly cleared due to the strong immunogenicity of bacterial-derived plasmid DNA, the low efficiency of Cre, and the bidirectional uninterrupted recombination, resulting in reduced stability of cccDNA in vivo, and HBV cannot Perform secondary infection, cannot simulate the entire life cycle of HBV infection

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