The Self-Inactivating KamiCas9 System for the Editing of CNS Disease Genes
Nicolas Merienne,
Gabriel Vachey,
Lucie de Longprez,
Cécile Meunier,
Virginie Zimmer,
Guillaume Perriard,
Mathieu Canales,
Amandine Mathias,
Lucas Herrgott,
Tim Beltraminelli,
Axelle Maulet,
Thomas Dequesne,
Catherine Pythoud,
Maria Rey,
Luc Pellerin,
Emmanuel Brouillet,
Anselme L. Perrier,
Renaud du Pasquier,
Nicole Déglon
Affiliations
Nicolas Merienne
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Gabriel Vachey
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Lucie de Longprez
CEA, DRF, Institut François Jacob, Molecular Imaging Research Center (MIRCen), F-92265 Fontenay-aux-Roses, France
Cécile Meunier
Department of Physiology, Laboratory of Neuroenergetics, University of Lausanne, 1011 Lausanne, Switzerland
Virginie Zimmer
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Guillaume Perriard
Department of Clinical Neurosciences, Laboratory of Neuro-immunology, Lausanne University Hospital, 1011 Lausanne, Switzerland
Mathieu Canales
Department of Clinical Neurosciences, Laboratory of Neuro-immunology, Lausanne University Hospital, 1011 Lausanne, Switzerland
Amandine Mathias
Department of Clinical Neurosciences, Laboratory of Neuro-immunology, Lausanne University Hospital, 1011 Lausanne, Switzerland
Lucas Herrgott
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Tim Beltraminelli
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Axelle Maulet
Institut National de la Santé et de la Recherche Médicale (INSERM) UMR861, I-Stem, AFM, 91100 Corbeil-Essonnes, France
Thomas Dequesne
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Catherine Pythoud
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Maria Rey
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Luc Pellerin
Department of Physiology, Laboratory of Neuroenergetics, University of Lausanne, 1011 Lausanne, Switzerland
Emmanuel Brouillet
CEA, DRF, Institut François Jacob, Molecular Imaging Research Center (MIRCen), F-92265 Fontenay-aux-Roses, France
Anselme L. Perrier
Institut National de la Santé et de la Recherche Médicale (INSERM) UMR861, I-Stem, AFM, 91100 Corbeil-Essonnes, France
Renaud du Pasquier
Department of Clinical Neurosciences, Laboratory of Neuro-immunology, Lausanne University Hospital, 1011 Lausanne, Switzerland
Nicole Déglon
Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland
Neurodegenerative disorders are a major public health problem because of the high frequency of these diseases. Genome editing with the CRISPR/Cas9 system is making it possible to modify the sequence of genes linked to these disorders. We designed the KamiCas9 self-inactivating editing system to achieve transient expression of the Cas9 protein and high editing efficiency. In the first application, the gene responsible for Huntington’s disease (HD) was targeted in adult mouse neuronal and glial cells. Mutant huntingtin (HTT) was efficiently inactivated in mouse models of HD, leading to an improvement in key markers of the disease. Sequencing of potential off-targets with the constitutive Cas9 system in differentiated human iPSC revealed a very low incidence with only one site above background level. This off-target frequency was significantly reduced with the KamiCas9 system. These results demonstrate the potential of the self-inactivating CRISPR/Cas9 editing for applications in the context of neurodegenerative diseases.
