Generation of a mouse model for the in vivo study of HCV-host interaction

D'Angelo, Fulvio (2010) Generation of a mouse model for the in vivo study of HCV-host interaction. [Tesi di dottorato] (Inedito)

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Abstract

The natural species tropism of hepatitis C virus (HCV) is limited to humans and higher primates. Due to the limited availability of chimpanzees, the establishment of a small-animal model for the study of HCV infection has a high priority. The recent identification of the human cellular receptors that are essential for HCV entry together with the advances in mouse genome engineering provides the bases for creating an inbred mouse model of HCV infection and propagation. In our study we designed a gene targeting strategy to produce novel humanized mouse strains expressing human-specific HCV entry receptors. Human genes for HCV essential entry factors were introduced in the mouse genome by homologous recombination in murine embryonic stem cells. To achieve the native expression patterns of their murine orthologues, we adopted a knock-in approach replacing the murine genes with human coding sequences. Three novel humanized mouse strains were generated, expressing respectively human CD81, scavenger receptor class B member 1 and occludin. Molecular analyses of mice genomic DNA and messenger RNA confirmed the presence and germ line propagation of the human genes, and their transcription. The native regulated expression of human-specific HCV entry receptors can overcome the major block to HCV replication in murine cells. To verify the virus entry into the cells of our humanized mice, we successfully optimized isolation and culture of murine primary hepatocytes and HCV pseudotype particles infection assay. Genetic humanization of mice for species-specific HCV receptors could allow HCV glycoproteins mediated cell entry and could be an important tool for in vivo studying of virus-host interactions.

Tipologia di documento:Tesi di dottorato
Parole chiave:Hepatitis C; mouse model
Settori scientifico-disciplinari MIUR:Area 06 Scienze mediche > MED/03 GENETICA MEDICA
Area 05 Scienze biologiche > BIO/18 GENETICA
Coordinatori della Scuola di dottorato:
Coordinatore del Corso di dottoratoe-mail (se nota)
Nitsch, Lucionitsch@unina.it
Tutor della Scuola di dottorato:
Tutor del Corso di dottoratoe-mail (se nota)
De Felice, Mario
Stato del full text:Accessibile
Data:29 Novembre 2010
Numero di pagine:71
Istituzione:Università degli studi di Napoli Federico II
Dipartimento o Struttura:Biologia e patologia cellullare e molecolare "L. Califano"
Tipo di tesi:Dottorato
Stato dell'Eprint:Inedito
Scuola di dottorato:Medicina molecolare
Denominazione del dottorato:Genetica e medicina molecolare
Ciclo di dottorato:22
Numero di sistema:8179
Depositato il:13 Dicembre 2010 23:30
Ultima modifica:29 Agosto 2012 12:21

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