title: Variable correction of Artemis deficiency by I-Sce1-meganuclease-assisted homologous recombination in murine hematopoietic stem cells.
abstract: The correction of genetic mutations by homologous recombination is an attractive approach to gene therapy. We used the DNA double-strand breaks introduced by the site-specific endonuclease I-Sce1 as a means of increasing homologous recombination of an exogenous DNA template in murine hematopoietic stem cells (mHSCs). To develop this approach, we chose an Artemis knockout (Art(-/-)) mouse in which exon 12 of the Artemis gene had been replaced by an I-Sce1 recognition site. The I-Sce1 enzyme and the Artemis correction template were each delivered by a self-inactivating (SIN)-integrase-defective lentiviral vector (SIN-IDLV-CMV-ISce1 and SIN-IDLV-Art, respectively). Transduction of Art(-/-) mHSCs with the two vectors successfully reverted the Art(-/-) phenotype in 2 of our 10 experiments. Even though the potential for genotoxicity has yet to be evaluated, this new approach to gene editing appears to be promising. Improving the efficacy of this approach will require further technical work.
authors.affiliation: 1] INSERM U768 Necker Hospital, Paris, France [2] IMAGINE Institute, Paris Descartes University, Paris, France.
authors.forename: J
authors.initials: J
authors.lastname: Rivière
authors.affiliation: 1] INSERM U768 Necker Hospital, Paris, France [2] Department of Pediatric Hematology and Oncology, Children's University Hospital, Düsseldorf, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Hauer
authors.affiliation: Cellectis Therapeutics, Paris, France.
authors.forename: L
authors.initials: L
authors.lastname: Poirot
authors.affiliation: 1] INSERM U768 Necker Hospital, Paris, France [2] IMAGINE Institute, Paris Descartes University, Paris, France.
authors.forename: J
authors.initials: J
authors.lastname: Brochet
authors.affiliation: Molecular Virology and Vectorology Laboratory, Pasteur Institute, Paris, France.
authors.forename: P
authors.initials: P
authors.lastname: Souque
authors.affiliation: Molecular Virology and Vectorology Laboratory, Pasteur Institute, Paris, France.
authors.forename: K
authors.initials: K
authors.lastname: Mollier
authors.affiliation: Cellectis Therapeutics, Paris, France.
authors.forename: A
authors.initials: A
authors.lastname: Gouble
authors.affiliation: Molecular Virology and Vectorology Laboratory, Pasteur Institute, Paris, France.
authors.forename: P
authors.initials: P
authors.lastname: Charneau
authors.affiliation: 1] INSERM U768 Necker Hospital, Paris, France [2] IMAGINE Institute, Paris Descartes University, Paris, France [3] Unit of Pediatric Immuno-Hematology, APHP, Necker Hospital, Paris, France.
authors.forename: A
authors.initials: A
authors.lastname: Fischer
authors.affiliation: Cellectis Therapeutics, Paris, France.
authors.forename: F
authors.initials: F
authors.lastname: Pâques
authors.affiliation: 1] INSERM U768 Necker Hospital, Paris, France [2] IMAGINE Institute, Paris Descartes University, Paris, France.
authors.forename: J-P
authors.initials: JP
authors.lastname: de Villartay
authors.affiliation: 1] INSERM U768 Necker Hospital, Paris, France [2] IMAGINE Institute, Paris Descartes University, Paris, France [3] Department of Biotherapy, APHP, Necker Hospital, Paris, France [4] CIC Biotherapy GHU West, INSERM-APHP, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Cavazzana
mesh_terms: D000818:Animals; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D015252:Deoxyribonucleases, Type II Site-Specific; D004720:Endonucleases / Q000172:deficiency / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D059765:Homologous Recombination / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D009687:Nuclear Proteins / Q000172:deficiency / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D014161:Transduction, Genetic
chemical_list: D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D004720:Endonucleases; C473337:Dclre1c protein, mouse; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific