Spell Checking Nature: Versatility of CRISPR/Cas9 for Developing Treatments for Inherited Disorders

Daria Wojtal; Dwi U Kemaladewi; Zeenat Malam; Sarah Abdullah; Tatianna W Y Wong; Elzbieta Hyatt; Zahra Baghestani; Sergio Pereira; James Stavropoulos; Vincent Mouly; Kamel Mamchaoui; Francesco Muntoni; Thomas Voit; Hernan D Gonorazky; James J Dowling; Michael D Wilson; Roberto Mendoza-Londono; Evgueni A Ivakine; Ronald D Cohn

doi:10.1016/j.ajhg.2015.11.012

Spell Checking Nature: Versatility of CRISPR/Cas9 for Developing Treatments for Inherited Disorders

Am J Hum Genet. 2016 Jan 7;98(1):90-101. doi: 10.1016/j.ajhg.2015.11.012. Epub 2015 Dec 10.

Authors

Daria Wojtal¹, Dwi U Kemaladewi², Zeenat Malam², Sarah Abdullah², Tatianna W Y Wong², Elzbieta Hyatt², Zahra Baghestani², Sergio Pereira³, James Stavropoulos⁴, Vincent Mouly⁵, Kamel Mamchaoui⁵, Francesco Muntoni⁶, Thomas Voit⁷, Hernan D Gonorazky⁸, James J Dowling⁹, Michael D Wilson¹, Roberto Mendoza-Londono¹⁰, Evgueni A Ivakine², Ronald D Cohn¹¹

Affiliations

¹ 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.
² Program in Genetics and Genome Biology, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 0A4, Canada.
³ 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.
⁴ Department of Paediatrics, University of Toronto, Toronto, ON M5G 1X8, Canada.
⁵ 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.
⁶ Dubowitz Neuromuscular Centre, Institute of Child Health and Great Ormond Street Hospital, London WC1N 1EH, UK.
⁷ NIHR Biomedical Research Centre, Institute of Child Health, University College London, 30 Guilford Street, London WC1N 1EH, UK.
⁸ 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.
⁹ 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.
¹⁰ 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.
¹¹ 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.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alleles
Clustered Regularly Interspaced Short Palindromic Repeats*
Gene Expression
Genetic Diseases, Inborn / genetics
Genetic Diseases, Inborn / therapy*
Humans
Muscular Dystrophy, Duchenne / genetics
Muscular Dystrophy, Duchenne / therapy