The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells
Lentiviral vectors designed for the treatment of the hemoglobinopathies require the inclusion of regulatory and strong enhancer elements to achieve sufficient expression of the β-globin transgene. Despite the inclusion of these elements, the efficacy of these vectors may be limited by transgene sil...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-01-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050116300249 |
| _version_ | 1819094875219427328 |
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| author | Zulema Romero Beatriz Campo-Fernandez Jennifer Wherley Michael L Kaufman Fabrizia Urbinati Aaron R Cooper Megan D Hoban Kismet M Baldwin Dianne Lumaquin Xiaoyan Wang Shantha Senadheera Roger P Hollis Donald B Kohn |
| author_facet | Zulema Romero Beatriz Campo-Fernandez Jennifer Wherley Michael L Kaufman Fabrizia Urbinati Aaron R Cooper Megan D Hoban Kismet M Baldwin Dianne Lumaquin Xiaoyan Wang Shantha Senadheera Roger P Hollis Donald B Kohn |
| author_sort | Zulema Romero |
| collection | DOAJ |
| description | Lentiviral vectors designed for the treatment of the hemoglobinopathies require the inclusion of regulatory and strong enhancer elements to achieve sufficient expression of the β-globin transgene. Despite the inclusion of these elements, the efficacy of these vectors may be limited by transgene silencing due to the genomic environment surrounding the integration site. Barrier insulators can be used to give more consistent expression and resist silencing even with lower vector copies. Here, the barrier activity of an insulator element from the human ankyrin-1 gene was analyzed in a lentiviral vector carrying an antisickling human β-globin gene. Inclusion of a single copy of the Ankyrin insulator did not affect viral titer, and improved the consistency of expression from the vector in murine erythroleukemia cells. The presence of the Ankyrin insulator element did not change transgene expression in human hematopoietic cells in short-term erythroid culture or in vivo in primary murine transplants. However, analysis in secondary recipients showed that the lentiviral vector with the Ankyrin element preserved transgene expression, whereas expression from the vector lacking the Ankyrin insulator decreased in secondary recipients. These studies demonstrate that the Ankyrin insulator may improve long-term β-globin expression in hematopoietic stem cells for gene therapy of hemoglobinopathies. |
| first_indexed | 2024-12-21T23:34:21Z |
| format | Article |
| id | doaj.art-af8dd3374c2f4e9592271ef1374da280 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-21T23:34:21Z |
| publishDate | 2015-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-af8dd3374c2f4e9592271ef1374da2802022-12-21T18:46:25ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012015-01-012The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cellsZulema Romero0Beatriz Campo-Fernandez1Jennifer Wherley2Michael L Kaufman3Fabrizia Urbinati4Aaron R Cooper5Megan D Hoban6Kismet M Baldwin7Dianne Lumaquin8Xiaoyan Wang9Shantha Senadheera10Roger P Hollis11Donald B Kohn12Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USAMolecular Biology Interdepartmental PhD Program, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Internal Medicine and Health Services Research, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USADepartment of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California, USA; Department of Pediatrics, UCLA Children's Discovery and Innovation Institute David Geffen School of Medicine, University of California, Los Angeles, California, USALentiviral vectors designed for the treatment of the hemoglobinopathies require the inclusion of regulatory and strong enhancer elements to achieve sufficient expression of the β-globin transgene. Despite the inclusion of these elements, the efficacy of these vectors may be limited by transgene silencing due to the genomic environment surrounding the integration site. Barrier insulators can be used to give more consistent expression and resist silencing even with lower vector copies. Here, the barrier activity of an insulator element from the human ankyrin-1 gene was analyzed in a lentiviral vector carrying an antisickling human β-globin gene. Inclusion of a single copy of the Ankyrin insulator did not affect viral titer, and improved the consistency of expression from the vector in murine erythroleukemia cells. The presence of the Ankyrin insulator element did not change transgene expression in human hematopoietic cells in short-term erythroid culture or in vivo in primary murine transplants. However, analysis in secondary recipients showed that the lentiviral vector with the Ankyrin element preserved transgene expression, whereas expression from the vector lacking the Ankyrin insulator decreased in secondary recipients. These studies demonstrate that the Ankyrin insulator may improve long-term β-globin expression in hematopoietic stem cells for gene therapy of hemoglobinopathies.http://www.sciencedirect.com/science/article/pii/S2329050116300249 |
| spellingShingle | Zulema Romero Beatriz Campo-Fernandez Jennifer Wherley Michael L Kaufman Fabrizia Urbinati Aaron R Cooper Megan D Hoban Kismet M Baldwin Dianne Lumaquin Xiaoyan Wang Shantha Senadheera Roger P Hollis Donald B Kohn The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells Molecular Therapy: Methods & Clinical Development |
| title | The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells |
| title_full | The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells |
| title_fullStr | The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells |
| title_full_unstemmed | The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells |
| title_short | The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells |
| title_sort | human ankyrin 1 promoter insulator sustains gene expression in a i² globin lentiviral vector in hematopoietic stem cells |
| url | http://www.sciencedirect.com/science/article/pii/S2329050116300249 |
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