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The first pooled, genome-wide CRISPR-Cas9 knockout (KO) screen with authentic HBV infection is performed and these results are integrated with two orthogonal pooled screens to identify host factors to expand the catalog of HBV host factors and provide a scalable framework for host target discovery.
Lipofection- and lentivirus-mediated protocols for CRISPR-Cas9 delivery in mouse-passaged primary human hepatocytes (mpPHH) are reported, a system that enables PHH expansion in liver-humanized mice and enables scalable genetic manipulation of mpPHH, opening new avenues for HBV research and liver disease modeling.
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