{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T20:55:44Z","timestamp":1790456144908,"version":"4.1.0"},"posted":{"date-parts":[[2026,7,24]]},"group-title":"Microbiology","reference-count":60,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T00:00:00Z","timestamp":1784851200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R21AI176944"],"award-info":[{"award-number":["R21AI176944"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R21AI176944-S1"],"award-info":[{"award-number":["R21AI176944-S1"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R01AI181682"],"award-info":[{"award-number":["R01AI181682"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R56AI182395"],"award-info":[{"award-number":["R56AI182395"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["DP1DA066168"],"award-info":[{"award-number":["DP1DA066168"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["DP1DK139804"],"award-info":[{"award-number":["DP1DK139804"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"award":["DRG-2440-21"],"award-info":[{"award-number":["DRG-2440-21"]}],"id":[{"id":"https:\/\/ror.org\/01gd7b947","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,24]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Hepatitis B virus (HBV) chronically infects approximately 250 million people worldwide, and reliable curative therapies are lacking. A broader understanding of viral-host interactions could accelerate efforts to find new host-centric therapeutic targets. However, inefficient cell culture systems and limited replication markers compatible with pooled screening have precluded the widespread use of genetic perturbation screens. Here, we performed the first pooled, genome-wide CRISPR-Cas9 knockout (KO) screen with authentic HBV infection and integrated these results with two orthogonal pooled screens to identify host factors. We selected 72 genes for a multi-step assessment that included arrayed validation assays using both HBV infection and pgRNA transfection. We then independently tested thirteen genes using high-efficiency bulk KO experiments to guide further investigations of both antiviral and proviral factors. In both KO and siRNA-mediated knockdown experiments, depletion of the top antiviral factor,\n                  <jats:italic>EXOC1<\/jats:italic>\n                  , enhanced multiple HBV replication markers, and transcriptomic analysis revealed activation of hypoxia- and HIF-1 gene signatures. Three proviral factors,\n                  <jats:italic>IRF2<\/jats:italic>\n                  ,\n                  <jats:italic>WDR48<\/jats:italic>\n                  , and\n                  <jats:italic>ZCCHC14<\/jats:italic>\n                  , were investigated\n                  <jats:italic>in vivo<\/jats:italic>\n                  using a human liver chimeric mouse model, which demonstrated that\n                  <jats:italic>ZCCHC14<\/jats:italic>\n                  KO greatly reduced HBV replication and spread. Together, these complementary\n                  <jats:italic>in vitro<\/jats:italic>\n                  and\n                  <jats:italic>in vivo<\/jats:italic>\n                  platforms expand the catalog of HBV host factors and provide a scalable framework for host target discovery.\n                <\/jats:p>","DOI":"10.64898\/2026.07.23.740386","type":"posted-content","created":{"date-parts":[[2026,7,25]],"date-time":"2026-07-25T00:11:17Z","timestamp":1784938277000},"source":"Crossref","is-referenced-by-count":1,"title":["Orthogonal CRISPR screens and human liver chimeric mice identify hepatitis B virus host factors"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1939-3102","authenticated-orcid":false,"given":"Catherine A","family":"Freije","sequence":"first","affiliation":[{"name":"Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY 10065, 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