Establishment Of The Transgenic Mice Which Express The HBV Receptor Human ASGPR

Hepatitis B is an infectious illness caused by hepatitis B virus(HBV),which causes an inflammation of liver and organs damage. Mice is a common animal model for human diseases, but mice can not be infected by HBV due to the strong species-specificity of HBV infection, its hosts are limited to minority primates,such as humans and chimpanzees, which resulted the shortage of appropriate susceptible animal model for the research of the pathogenic mechanism; or the clinical test of drugs and vaccines. The human asialoglycoprotein receptor (ASGPR) is a transmembrane molecule specifically expressed on the sinusoidal and basolateral hepatocellular membrane mediating uptake and intracellular degradation of desialylated glycoproteins by liver cells. It is constructed of two subunits of related amino acid sequence, H1 and H2. Expression of both H1and H2 cDNAs is required to generate high affinity asialoglycoprotein binding sites.A study suggests that the hepatic ASGPR is able to bind natural HBV specifically with its preS1 region and that this attachment might provide a mechanism for the hepatic endocytosis of HBV.Our research use the microinjection technique to generate the transgenic mice which express the HBV receptor human ASGPR. Our study will provide HBV susceptible animal models for the relevant research.Methods:Both cDNA of the Human ASGPR H1&H2 subunits were cloned and the expression vector PCAGGS-ASGPRH1&H2 were constructed, after the linearization by SalI and DrdI, Both of the 3.9kb DNA fragments were introduced into the fertilized eggs of mice by microinjection;The genotypes of the transgenic mice were identified by PCR and Southern-blot methods;The transcription of the transgene were identified by RT-PCR method;The expression of the exogenous ASGPR H1 and ASGPR H2 genes were confirmed by Western-blot.Results: 5 Founfers which integrate both the ASGPR H1 and ASGPR H2 genes were generated. The positive rate of co-microinjection is 9.6%. Transcription of ASGPR H1 and ASGPR H2 were detected in some of the F1 generation mice.One transgenic mouse line that express the human ASGPR was obtained. Conclusion: The transgenic mouse line was establsihed which express the HBV receptor human ASGPR in the liver tissue of transgenic mice.This study is hoped to provide a good animal infection model for HBV research.