CRISPR-mediated rapid generation of neural cell-specific knockout mice facilitates research in neurophysiology and pathology
Inducible conditional knockout mice are important tools for studying gene function and disease therapy, but their generation is costly and time-consuming. We introduced clustered regularly interspaced short palindromic repeats (CRISPR) and Cre into an LSL-Cas9 transgene-carrying mouse line by using...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-03-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050121000279 |
| _version_ | 1818610200861474816 |
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| author | Dan Xiao Weifeng Zhang Qing Wang Xing Li Yuan Zhang Javad Rasouli Giacomo Casella Bogoljub Ciric Mark Curtis Abdolmohamad Rostami Guang-Xian Zhang |
| author_facet | Dan Xiao Weifeng Zhang Qing Wang Xing Li Yuan Zhang Javad Rasouli Giacomo Casella Bogoljub Ciric Mark Curtis Abdolmohamad Rostami Guang-Xian Zhang |
| author_sort | Dan Xiao |
| collection | DOAJ |
| description | Inducible conditional knockout mice are important tools for studying gene function and disease therapy, but their generation is costly and time-consuming. We introduced clustered regularly interspaced short palindromic repeats (CRISPR) and Cre into an LSL-Cas9 transgene-carrying mouse line by using adeno-associated virus (AAV)-PHP.eB to rapidly knockout gene(s) specifically in central nervous system (CNS) cells of adult mice. NeuN in neurons and GFAP in astrocytes were knocked out 2 weeks after an intravenous injection of vector, with an efficiency comparable to that of inducible Cre-loxP conditional knockout. For functional testing, we generated astrocyte-specific Act1 knockout mice, which exhibited a phenotype similar to mice with Cre-loxP-mediated Act1 knockout, in an animal model of multiple sclerosis (MS), an autoimmune disorder of the CNS. With this novel technique, neural cell-specific knockout can be induced rapidly (few weeks) and cost-effectively. Our study provides a new approach to building inducible conditional knockout mice, which would greatly facilitate research on CNS biology and disease. |
| first_indexed | 2024-12-16T15:10:40Z |
| format | Article |
| id | doaj.art-6866c8fcb81e45ca835208c0278dd192 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-16T15:10:40Z |
| publishDate | 2021-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-6866c8fcb81e45ca835208c0278dd1922022-12-21T22:26:58ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012021-03-0120755764CRISPR-mediated rapid generation of neural cell-specific knockout mice facilitates research in neurophysiology and pathologyDan Xiao0Weifeng Zhang1Qing Wang2Xing Li3Yuan Zhang4Javad Rasouli5Giacomo Casella6Bogoljub Ciric7Mark Curtis8Abdolmohamad Rostami9Guang-Xian Zhang10Department of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USADepartment of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USA; College of Life Sciences, Shaanxi Normal University, Xi’an, Shaanxi 710062, ChinaDepartment of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USACollege of Life Sciences, Shaanxi Normal University, Xi’an, Shaanxi 710062, ChinaCollege of Life Sciences, Shaanxi Normal University, Xi’an, Shaanxi 710062, ChinaDepartment of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USADepartment of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USADepartment of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USADepartment of Pathology, Thomas Jefferson University, Philadelphia, PA 19107, USADepartment of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USADepartment of Neurology, Thomas Jefferson University, Philadelphia, PA 19107, USA; Corresponding author: Guang-Xian Zhang, Department of Neurology, Thomas Jefferson University, JHN 300, Philadelphia, PA 19107, USA.Inducible conditional knockout mice are important tools for studying gene function and disease therapy, but their generation is costly and time-consuming. We introduced clustered regularly interspaced short palindromic repeats (CRISPR) and Cre into an LSL-Cas9 transgene-carrying mouse line by using adeno-associated virus (AAV)-PHP.eB to rapidly knockout gene(s) specifically in central nervous system (CNS) cells of adult mice. NeuN in neurons and GFAP in astrocytes were knocked out 2 weeks after an intravenous injection of vector, with an efficiency comparable to that of inducible Cre-loxP conditional knockout. For functional testing, we generated astrocyte-specific Act1 knockout mice, which exhibited a phenotype similar to mice with Cre-loxP-mediated Act1 knockout, in an animal model of multiple sclerosis (MS), an autoimmune disorder of the CNS. With this novel technique, neural cell-specific knockout can be induced rapidly (few weeks) and cost-effectively. Our study provides a new approach to building inducible conditional knockout mice, which would greatly facilitate research on CNS biology and disease.http://www.sciencedirect.com/science/article/pii/S2329050121000279CRISPRConditional knockoutAAV PHP.eBNeural-specific knockoutMultiple sclerosisKnockout mouse model |
| spellingShingle | Dan Xiao Weifeng Zhang Qing Wang Xing Li Yuan Zhang Javad Rasouli Giacomo Casella Bogoljub Ciric Mark Curtis Abdolmohamad Rostami Guang-Xian Zhang CRISPR-mediated rapid generation of neural cell-specific knockout mice facilitates research in neurophysiology and pathology Molecular Therapy: Methods & Clinical Development CRISPR Conditional knockout AAV PHP.eB Neural-specific knockout Multiple sclerosis Knockout mouse model |
| title | CRISPR-mediated rapid generation of neural cell-specific knockout mice facilitates research in neurophysiology and pathology |
| title_full | CRISPR-mediated rapid generation of neural cell-specific knockout mice facilitates research in neurophysiology and pathology |
| title_fullStr | CRISPR-mediated rapid generation of neural cell-specific knockout mice facilitates research in neurophysiology and pathology |
| title_full_unstemmed | CRISPR-mediated rapid generation of neural cell-specific knockout mice facilitates research in neurophysiology and pathology |
| title_short | CRISPR-mediated rapid generation of neural cell-specific knockout mice facilitates research in neurophysiology and pathology |
| title_sort | crispr mediated rapid generation of neural cell specific knockout mice facilitates research in neurophysiology and pathology |
| topic | CRISPR Conditional knockout AAV PHP.eB Neural-specific knockout Multiple sclerosis Knockout mouse model |
| url | http://www.sciencedirect.com/science/article/pii/S2329050121000279 |
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