title: Inhibition of JCPyV infection mediated by targeted viral genome editing using CRISPR/Cas9.
abstract: Progressive multifocal leukoencephalopathy (PML) is a debilitating disease resulting from infection of oligodendrocytes by the JC polyomavirus (JCPyV). Currently, there is no anti-viral therapeutic available against JCPyV infection. The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system (CRISPR/Cas9) is a genome editing tool capable of introducing sequence specific breaks in double stranded DNA. Here we show that the CRISPR/Cas9 system can restrict the JCPyV life cycle in cultured cells. We utilized CRISPR/Cas9 to target the noncoding control region and the late gene open reading frame of the JCPyV genome. We found significant inhibition of virus replication and viral protein expression in cells recipient of Cas9 together with JCPyV-specific single-guide RNA delivered prior to or after JCPyV infection.
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Yi-Ying
authors.initials: YY
authors.lastname: Chou
authors.affiliation: Department of Neuroimmunology, Biogen, Cambridge, MA 02142, USA.
authors.forename: Annabel
authors.initials: A
authors.lastname: Krupp
authors.affiliation: Department of Neuroimmunology, Biogen, Cambridge, MA 02142, USA.
authors.forename: Campbell
authors.initials: C
authors.lastname: Kaynor
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Gaudin
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Minghe
authors.initials: M
authors.lastname: Ma
authors.affiliation: Department of Neuroimmunology, Biogen, Cambridge, MA 02142, USA.
authors.forename: Ellen
authors.initials: E
authors.lastname: Cahir-McFarland
authors.affiliation: Department of Cell Biology, Harvard Medical School, and Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115, USA.|Department of Pediatrics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Tom
authors.initials: T
authors.lastname: Kirchhausen
mesh_terms: D064113:CRISPR-Cas Systems; D000072669:Gene Editing / Q000379:methods*; D016679:Genome, Viral / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D007577:JC Virus / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D007968:Leukoencephalopathy, Progressive Multifocal / Q000821:virology*; D016366:Open Reading Frames / Q000187:drug effects; D027601:Polyomavirus Infections / Q000821:virology*; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000494:pharmacology; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D014764:Viral Proteins