1. The Self-Inactivating KamiCas9 System for the Editing of CNS Disease Genes
- Author
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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, and Nicole Déglon
- Subjects
Cerebral Cortex ,Gene Editing ,Neurons ,Huntingtin Protein ,Base Sequence ,Induced Pluripotent Stem Cells ,lentiviral vectors ,self-inactivating system ,Kinetics ,Mice ,KamiCas9 ,HEK293 Cells ,lcsh:Biology (General) ,Central Nervous System Diseases ,Astrocytes ,Animals ,Astrocytes/cytology ,Astrocytes/metabolism ,CRISPR-Cas Systems/genetics ,Cells, Cultured ,Central Nervous System Diseases/genetics ,Cerebral Cortex/cytology ,Humans ,Huntingtin Protein/genetics ,Induced Pluripotent Stem Cells/cytology ,Induced Pluripotent Stem Cells/metabolism ,Neurons/cytology ,Neurons/metabolism ,CRISPR/Cas9 ,Huntington’s disease ,gene editing ,neurodegenerative diseases ,CRISPR-Cas Systems ,lcsh:QH301-705.5 - Abstract
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.
- Published
- 2016