Cd302 and Cr1l restrict human hepatotropic virus cross-species transmission to mice [6PMH siRNA]

Virus species- and tissue-tropism is governed by host dependency and restriction factors. Hepatitis C virus (HCV) exhibits a narrow species-tropism and murine hepatocytes are refractory to infection. Using murine liver cDNA library screening we identified Cd302, a lectin, and Cr1l, a complement receptor, as pan-genotypic restrictors of HCV infection. Cd302/Cr1l interact to impede virion uptake and co-operatively induce a non-canonical transcriptional program, inhibiting HCV and hepatitis B virus (HBV) infection in vitro. CAS9 disruption of murine hepatocyte Cd302 expression increased HCV permissiveness in-vivo and ex-vivo, and modulated the intrinsic hepatocyte transcriptome dysregulating metabolic process and host defense genes. In contrast, co-operative CD302/CR1L expression was absent and HCV restriction reduced in human hepatocytes. The Cd302/Cr1l axis therefore contributes to limiting hepatotropic virus cross-species transmission to mice, opening new avenues for step-wise development of mouse models for these important human pathogens, which cause substantial disease burden globally. Primary mouse hepatocytes from x3 entry receptor transgenic mice with conditional liver expression (human Occludin and CD81: hOChep) and one wildtype control mouse were isolated after liver perfusion and liberase digestion. Hepatoyctes were plated onto 6-well tissue culture dishes dishes and cultured in HCM. 24 hours after plating hepatocytes were transfected with siRNAs targeting the genes of interest or a control siRNA. 24 hours after siRNA transfection hepatocytes were infected with HCV (strain Jc1, MOI 1) or left uninfected. 24 hours post infection RNA was isolated for RNA-seq.