title: Genome-wide inactivation of porcine endogenous retroviruses (PERVs).
abstract: The shortage of organs for transplantation is a major barrier to the treatment of organ failure. Although porcine organs are considered promising, their use has been checked by concerns about the transmission of porcine endogenous retroviruses (PERVs) to humans. Here we describe the eradication of all PERVs in a porcine kidney epithelial cell line (PK15). We first determined the PK15 PERV copy number to be 62. Using CRISPR-Cas9, we disrupted all copies of the PERV pol gene and demonstrated a >1000-fold reduction in PERV transmission to human cells, using our engineered cells. Our study shows that CRISPR-Cas9 multiplexability can be as high as 62 and demonstrates the possibility that PERVs can be inactivated for clinical application of porcine-to-human xenotransplantation.
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA. Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA. eGenesis Biosciences, Boston, MA 02115, USA. gchurch@genetics.med.harvard.edu luhan.yang@egenesisbio.com.
authors.forename: Luhan
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA. Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA. eGenesis Biosciences, Boston, MA 02115, USA.
authors.forename: Marc
authors.initials: M
authors.lastname: Güell
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA. College of Animal Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Dong
authors.initials: D
authors.lastname: Niu
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA.
authors.forename: Haydy
authors.initials: H
authors.lastname: George
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA.
authors.forename: Emal
authors.initials: E
authors.lastname: Lesha
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA.
authors.forename: Dennis
authors.initials: D
authors.lastname: Grishin
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA.
authors.forename: John
authors.initials: J
authors.lastname: Aach
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA.
authors.forename: Ellen
authors.initials: E
authors.lastname: Shrock
authors.affiliation: Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
authors.forename: Weihong
authors.initials: W
authors.lastname: Xu
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Poci
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA.
authors.forename: Rebeca
authors.initials: R
authors.lastname: Cortazio
authors.affiliation: Transplant Infectious Disease and Compromised Host Program, Massachusetts General Hospital, Boston, MA 02115, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Wilkinson
authors.affiliation: Transplant Infectious Disease and Compromised Host Program, Massachusetts General Hospital, Boston, MA 02115, USA.
authors.forename: Jay A
authors.initials: JA
authors.lastname: Fishman
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA, USA. Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA. eGenesis Biosciences, Boston, MA 02115, USA. gchurch@genetics.med.harvard.edu luhan.yang@egenesisbio.com.
authors.forename: George
authors.initials: G
authors.lastname: Church
mesh_terms: D000818:Animals; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D002460:Cell Line; D020077:Endogenous Retroviruses / Q000235:genetics*; D004847:Epithelial Cells / Q000821:virology; D018628:Gene Dosage; D018390:Gene Targeting / Q000379:methods*; D015751:Genes, pol; D057809:HEK293 Cells; D006801:Humans; D007668:Kidney / Q000821:virology; D008969:Molecular Sequence Data; D012192:Retroviridae Infections / Q000517:prevention & control* / Q000635:transmission / Q000821:virology; D013552:Swine / Q000821:virology*; D014183:Transplantation, Heterologous / Q000379:methods*; D038301:Virus Inactivation