A CRISPR Screen Identifies the Cell Polarity Determinant Crumbs 3 as an Adeno-associated Virus Restriction Factor in Hepatocytes

Journal of Virology
Victoria J MadiganAravind Asokan

Abstract

Adeno-associated viruses (AAV) are helper-dependent parvoviruses that have been developed into promising gene therapy vectors. Many studies, including a recent unbiased genomic screen, have identified host factors essential for AAV cell entry, but no genome-wide screens that address inhibitory host factors have been reported. Here, we utilize a novel CRISPR screen to identify AAV restriction factors in a human hepatocyte cell line. The major hit from our gain-of-function screen is the apical polarity determinant Crumbs 3 (Crb3). Knockout (KO) of Crb3 enhances AAV transduction, while overexpression exerts the opposite effect. Further, Crb3 appears to restrict AAV transduction in a serotype- and cell type-specific manner. Particularly, for AAV serotype 9 and a rationally engineered AAV variant, we demonstrate that increased availability of galactosylated glycans on the surfaces of Crb3 KO cells, but not the universal AAV receptor, leads to increased capsid attachment and enhanced transduction. We postulate that Crb3 could serve as a key molecular determinant that restricts the availability of AAV glycan attachment factors on the cell surface by maintaining apical-basal polarity and tight junction integrity.IMPORTANCE Adeno-associ...Continue Reading

References

Jun 18, 1997·Biochemical and Biophysical Research Communications·Y J KimY C Lee
Dec 6, 2001·Proceedings of the National Academy of Sciences of the United States of America·C J CohenJ M Bergelson
Jul 24, 2002·Proceedings of the National Academy of Sciences of the United States of America·Qing XieMichael S Chapman
Jan 14, 2004·Molecular Biology of the Cell·Céline LemmersAndré Le Bivic
Jun 25, 2005·Journal of Cell Science·Vanessa C FoggBen Margolis
Nov 23, 2005·Bioinformatics·Konstantin ArnoldTorsten Schwede
Dec 22, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Giovanni Di Pasquale, John A Chiorini
Oct 26, 2007·Journal of Virology·Christopher J MeeJane A McKeating
Nov 5, 2008·Nucleic Acids Research·Mauricio Carrillo-TrippVijay S Reddy
Apr 10, 2009·Journal of Virology·Christopher J MeeJane A McKeating
Apr 13, 2010·Acta Crystallographica. Section D, Biological Crystallography·P EmsleyK Cowtan
Feb 19, 2011·The Journal of Biological Chemistry·Shen ShenAravind Asokan
Apr 13, 2012·Journal of Virology·Michael A DiMattiaMavis Agbandje-McKenna
Aug 30, 2012·Nature Methods·Caroline A SchneiderKevin W Eliceiri
Feb 27, 2013·The Journal of Cell Biology·Junya HayaseHideki Sumimoto
May 31, 2013·Comprehensive Physiology·Aleksandr Treyer, Anne Müsch
Oct 30, 2013·Molecular and Cellular Biology·Eileen L WhitemanBen Margolis
Dec 18, 2013·Science·Ophir ShalemFeng Zhang
Jul 23, 2014·Current Opinion in Virology·Lin-Ya HuangMavis Agbandje-McKenna
Jul 31, 2014·Nature Methods·Neville E SanjanaFeng Zhang
Jun 28, 2015·Journal of Hepatology·Paul Gissen, Irwin M Arias
Aug 4, 2015·Developmental Cell·Aleksander D SzymaniakXaralabos Varelas
Aug 26, 2015·Proceedings of the National Academy of Sciences of the United States of America·Miguel ManoMauro Giacca
Dec 2, 2015·PLoS Pathogens·Christina HölscherMartin Müller
Jan 28, 2016·Nature·S PillayJ E Carette
May 7, 2016·Cellular and Molecular Life Sciences : CMLS·Peter Coopman, Alexandre Djiane
Jun 5, 2016·Current Opinion in Virology·Victoria J Madigan, Aravind Asokan
Aug 22, 2016·Current Opinion in Virology·Garrett Edward Berry, Aravind Asokan
Sep 24, 2016·Molecular Cancer Research : MCR·Maroof AlamDonald Kufe
Aug 17, 2017·Cell Reports·Brook E HeatonNicholas S Heaton
Nov 28, 2017·Molecular Therapy : the Journal of the American Society of Gene Therapy·Blake H AlbrightAravind Asokan
Jan 19, 2018·Journal of Virology·Amanda M DudekLuk H Vandenberghe
Apr 10, 2019·Gene Therapy·Bradley A HamiltonJoseph Zabner

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Citations

Aug 9, 2020·Frontiers in Molecular Neuroscience·Andrew P Tosolini, James N Sleigh
Oct 20, 2020·Trends in Molecular Medicine·Bijay P DhungelJohn E J Rasko

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