The Self-Inactivating KamiCas9 System for the Editing of CNS Disease Genes

Cell Reports

Lucie de Longprez¹  Emmanuel Brouillet¹  Tim Beltraminelli²  Virginie Zimmer²  Nicolas Merienne²  Catherine Pythoud²  Gabriel Vachey²  Lucas Herrgott²  Nicole Déglon²  Maria Rey²  Thomas Dequesne²  Guillaume Perriard³  Amandine Mathias³  Mathieu Canales³  Renaud du Pasquier³  Cécile Meunier⁴  Luc Pellerin⁴  Anselme L. Perrier⁵  Axelle Maulet⁵ 

[1] CEA, DRF, Institut François Jacob, Molecular Imaging Research Center (MIRCen), F-92265 Fontenay-aux-Roses, France;Department of Clinical Neurosciences, Laboratory of Cellular and Molecular Neurotherapies (LCMN), Lausanne University Hospital, 1011 Lausanne, Switzerland;Department of Clinical Neurosciences, Laboratory of Neuro-immunology, Lausanne University Hospital, 1011 Lausanne, Switzerland;Department of Physiology, Laboratory of Neuroenergetics, University of Lausanne, 1011 Lausanne, Switzerland;Institut National de la Santé et de la Recherche Médicale (INSERM) UMR861, I-Stem, AFM, 91100 Corbeil-Essonnes, France;

关键词: CRISPR/Cas9;    gene editing;    self-inactivating system;    KamiCas9;    neurodegenerative diseases;    Huntington’s disease;    lentiviral vectors;   

DOI  :  10.1016/j.celrep.2017.08.075

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown