CRISPR-Cas9-Mediated Genome Editing Increases Lifespan and Improves Motor Deficits in a Huntington’s Disease Mouse Model
Huntington’s disease (HD) is a currently incurable and, ultimately, fatal neurodegenerative disorder caused by a CAG trinucleotide repeat expansion within exon 1 of the huntingtin (HTT) gene, which results in the production of a mutant protein that forms inclusions and selectively destroys neurons i...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2019-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253119301994 |
| _version_ | 1811325482002546688 |
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| author | Freja K. Ekman David S. Ojala Maroof M. Adil Paola A. Lopez David V. Schaffer Thomas Gaj |
| author_facet | Freja K. Ekman David S. Ojala Maroof M. Adil Paola A. Lopez David V. Schaffer Thomas Gaj |
| author_sort | Freja K. Ekman |
| collection | DOAJ |
| description | Huntington’s disease (HD) is a currently incurable and, ultimately, fatal neurodegenerative disorder caused by a CAG trinucleotide repeat expansion within exon 1 of the huntingtin (HTT) gene, which results in the production of a mutant protein that forms inclusions and selectively destroys neurons in the striatum and other adjacent structures. The RNA-guided Cas9 endonuclease from CRISPR-Cas9 systems is a versatile technology for inducing DNA double-strand breaks that can stimulate the introduction of frameshift-inducing mutations and permanently disable mutant gene function. Here, we show that the Cas9 nuclease from Staphylococcus aureus, a small Cas9 ortholog that can be packaged alongside a single guide RNA into a single adeno-associated virus (AAV) vector, can be used to disrupt the expression of the mutant HTT gene in the R6/2 mouse model of HD following its in vivo delivery to the striatum. Specifically, we found that CRISPR-Cas9-mediated disruption of the mutant HTT gene resulted in a ∼50% decrease in neuronal inclusions and significantly improved lifespan and certain motor deficits. These results thus illustrate the potential for CRISPR-Cas9 technology to treat HD and other autosomal dominant neurodegenerative disorders caused by a trinucleotide repeat expansion via in vivo genome editing. Keywords: CRISPR-Cas9, Huntington’s disease, AAV, gene therapy, genome editing |
| first_indexed | 2024-04-13T14:33:55Z |
| format | Article |
| id | doaj.art-9124d693316b4ce19eb68f74da73bd30 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-04-13T14:33:55Z |
| publishDate | 2019-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-9124d693316b4ce19eb68f74da73bd302022-12-22T02:43:05ZengElsevierMolecular Therapy: Nucleic Acids2162-25312019-09-0117829839CRISPR-Cas9-Mediated Genome Editing Increases Lifespan and Improves Motor Deficits in a Huntington’s Disease Mouse ModelFreja K. Ekman0David S. Ojala1Maroof M. Adil2Paola A. Lopez3David V. Schaffer4Thomas Gaj5Department of Chemistry, University of California, Berkeley, Berkeley, CA, USADepartment of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA, USADepartment of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA, USADepartment of Bioengineering, University of California, Berkeley, Berkeley, CA, USADepartment of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA, USA; Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA; Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA; The Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, CA, USA; Corresponding author: David V. Schaffer, Department of Chemical and Biomolecular Engineering, University of California, Berkeley, Berkeley, CA, USA.Department of Bioengineering, University of Illinois, Urbana, IL, USA; Carl R. Woese Institute for Genomic Biology, University of Illinois, Urbana, IL, USA; Corresponding author: Thomas Gaj, Department of Bioengineering, University of Illinois, Urbana, IL, USA.Huntington’s disease (HD) is a currently incurable and, ultimately, fatal neurodegenerative disorder caused by a CAG trinucleotide repeat expansion within exon 1 of the huntingtin (HTT) gene, which results in the production of a mutant protein that forms inclusions and selectively destroys neurons in the striatum and other adjacent structures. The RNA-guided Cas9 endonuclease from CRISPR-Cas9 systems is a versatile technology for inducing DNA double-strand breaks that can stimulate the introduction of frameshift-inducing mutations and permanently disable mutant gene function. Here, we show that the Cas9 nuclease from Staphylococcus aureus, a small Cas9 ortholog that can be packaged alongside a single guide RNA into a single adeno-associated virus (AAV) vector, can be used to disrupt the expression of the mutant HTT gene in the R6/2 mouse model of HD following its in vivo delivery to the striatum. Specifically, we found that CRISPR-Cas9-mediated disruption of the mutant HTT gene resulted in a ∼50% decrease in neuronal inclusions and significantly improved lifespan and certain motor deficits. These results thus illustrate the potential for CRISPR-Cas9 technology to treat HD and other autosomal dominant neurodegenerative disorders caused by a trinucleotide repeat expansion via in vivo genome editing. Keywords: CRISPR-Cas9, Huntington’s disease, AAV, gene therapy, genome editinghttp://www.sciencedirect.com/science/article/pii/S2162253119301994 |
| spellingShingle | Freja K. Ekman David S. Ojala Maroof M. Adil Paola A. Lopez David V. Schaffer Thomas Gaj CRISPR-Cas9-Mediated Genome Editing Increases Lifespan and Improves Motor Deficits in a Huntington’s Disease Mouse Model Molecular Therapy: Nucleic Acids |
| title | CRISPR-Cas9-Mediated Genome Editing Increases Lifespan and Improves Motor Deficits in a Huntington’s Disease Mouse Model |
| title_full | CRISPR-Cas9-Mediated Genome Editing Increases Lifespan and Improves Motor Deficits in a Huntington’s Disease Mouse Model |
| title_fullStr | CRISPR-Cas9-Mediated Genome Editing Increases Lifespan and Improves Motor Deficits in a Huntington’s Disease Mouse Model |
| title_full_unstemmed | CRISPR-Cas9-Mediated Genome Editing Increases Lifespan and Improves Motor Deficits in a Huntington’s Disease Mouse Model |
| title_short | CRISPR-Cas9-Mediated Genome Editing Increases Lifespan and Improves Motor Deficits in a Huntington’s Disease Mouse Model |
| title_sort | crispr cas9 mediated genome editing increases lifespan and improves motor deficits in a huntington s disease mouse model |
| url | http://www.sciencedirect.com/science/article/pii/S2162253119301994 |
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