title: Spell Checking Nature: Versatility of CRISPR/Cas9 for Developing Treatments for Inherited Disorders.
abstract: Clustered regularly interspaced short palindromic repeat (CRISPR) has arisen as a frontrunner for efficient genome engineering. However, the potentially broad therapeutic implications are largely unexplored. Here, to investigate the therapeutic potential of CRISPR/Cas9 in a diverse set of genetic disorders, we establish a pipeline that uses readily obtainable cells from affected individuals. We show that an adapted version of CRISPR/Cas9 increases the amount of utrophin, a known disease modifier in Duchenne muscular dystrophy (DMD). Furthermore, we demonstrate preferential elimination of the dominant-negative FGFR3 c.1138G>A allele in fibroblasts of an individual affected by achondroplasia. Using a previously undescribed approach involving single guide RNA, we successfully removed large genome rearrangement in primary cells of an individual with an X chromosome duplication including MECP2. Moreover, removal of a duplication of DMD exons 18-30 in myotubes of an individual affected by DMD produced full-length dystrophin. Our findings establish the far-reaching therapeutic utility of CRISPR/Cas9, which can be tailored to target numerous inherited disorders.
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.
authors.forename: Daria
authors.initials: D
authors.lastname: Wojtal
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
authors.forename: Dwi U
authors.initials: DU
authors.lastname: Kemaladewi
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
authors.forename: Zeenat
authors.initials: Z
authors.lastname: Malam
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
authors.forename: Sarah
authors.initials: S
authors.lastname: Abdullah
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
authors.forename: Tatianna W Y
authors.initials: TW
authors.lastname: Wong
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Hyatt
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Baghestani
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; The Centre for Applied Genomics, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
authors.forename: Sergio
authors.initials: S
authors.lastname: Pereira
authors.affiliation: Department of Paediatrics, University of Toronto, Toronto, ON M5G 1X8, Canada.
authors.forename: James
authors.initials: J
authors.lastname: Stavropoulos
authors.affiliation: INSERM UMRS974, Centre National de la Recherche Scientifique FRE3617, Center for Research in Myology, Université Pierre et Marie Curie (Paris 6), Sorbonne Universités, 47 Boulevard de l'Hôpital, 75013 Paris, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Mouly
authors.affiliation: INSERM UMRS974, Centre National de la Recherche Scientifique FRE3617, Center for Research in Myology, Université Pierre et Marie Curie (Paris 6), Sorbonne Universités, 47 Boulevard de l'Hôpital, 75013 Paris, France.
authors.forename: Kamel
authors.initials: K
authors.lastname: Mamchaoui
authors.affiliation: Dubowitz Neuromuscular Centre, Institute of Child Health and Great Ormond Street Hospital, London WC1N 1EH, UK.
authors.forename: Francesco
authors.initials: F
authors.lastname: Muntoni
authors.affiliation: NIHR Biomedical Research Centre, Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Voit
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Department of Paediatrics, University of Toronto, Toronto, ON M5G 1X8, Canada.
authors.forename: Hernan D
authors.initials: HD
authors.lastname: Gonorazky
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Paediatrics, University of Toronto, Toronto, ON M5G 1X8, Canada; Division of Clinical and Metabolic Genetics, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada.
authors.forename: James J
authors.initials: JJ
authors.lastname: Dowling
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Wilson
authors.affiliation: Department of Paediatrics, University of Toronto, Toronto, ON M5G 1X8, Canada; Division of Clinical and Metabolic Genetics, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada.
authors.forename: Roberto
authors.initials: R
authors.lastname: Mendoza-Londono
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
authors.forename: Evgueni A
authors.initials: EA
authors.lastname: Ivakine
authors.affiliation: Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Paediatrics, University of Toronto, Toronto, ON M5G 1X8, Canada; Division of Clinical and Metabolic Genetics, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada; Centre for Genetic Medicine, The Hospital for Sick Children, Toronto, ON M5G 1X8, Canada. Electronic address: ronald.cohn@sickkids.ca.
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Cohn
mesh_terms: D000483:Alleles; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D015870:Gene Expression; D030342:Genetic Diseases, Inborn / Q000235:genetics / Q000628:therapy*; D006801:Humans; D020388:Muscular Dystrophy, Duchenne / Q000235:genetics / Q000628:therapy