Repeatable, Inducible Micro-RNA-Based Technology Tightly Controls Liver Transgene Expression
Inducible systems for gene expression emerge as a new class of artificial vectors offering temporal and spatial exogenous control of gene expression. However, most inducible systems are less efficient in vivo and lack the target-organ specificity. In the present study, we have developed and optimize...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2014-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253116303146 |
| _version_ | 1819212560975527936 |
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| author | Iulian I Oprea Joana R Viola Pedro M D Moreno Oscar E Simonson Sergey Rodin Nathalie Teller Karl Tryggvason Karin E Lundin Leonard Girnita Carl Inge Edvard Smith |
| author_facet | Iulian I Oprea Joana R Viola Pedro M D Moreno Oscar E Simonson Sergey Rodin Nathalie Teller Karl Tryggvason Karin E Lundin Leonard Girnita Carl Inge Edvard Smith |
| author_sort | Iulian I Oprea |
| collection | DOAJ |
| description | Inducible systems for gene expression emerge as a new class of artificial vectors offering temporal and spatial exogenous control of gene expression. However, most inducible systems are less efficient in vivo and lack the target-organ specificity. In the present study, we have developed and optimized an oligonucleotide-based inducible system for the in vivo control of transgenes in the liver. We generated a set of simple, inducible plasmid-vectors based on the addition of four units of liver-specific miR-122 target sites to the 3′untranslated region of the gene of interest. Once the vector was delivered into hepatocytes this modification induced a dramatic reduction of gene expression that could be restored by the infusion of an antagomir for miR-122. The efficiency of the system was tested in vivo, and displayed low background and strong increase in gene expression upon induction. Moreover, gene expression was repeatedly induced even several months after the first induction showing no toxic effect in vivo. By combining tissue-specific control elements with antagomir treatment we generated, optimized and validated a robust inducible system that could be used successfully for in vivo experimental models requiring tight and cyclic control of gene expression. |
| first_indexed | 2024-12-23T06:44:55Z |
| format | Article |
| id | doaj.art-502466cfc7d34aeebe0b6b19531e8534 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-23T06:44:55Z |
| publishDate | 2014-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-502466cfc7d34aeebe0b6b19531e85342022-12-21T17:56:36ZengElsevierMolecular Therapy: Nucleic Acids2162-25312014-01-013C10.1038/mtna.2014.25Repeatable, Inducible Micro-RNA-Based Technology Tightly Controls Liver Transgene ExpressionIulian I Oprea0Joana R Viola1Pedro M D Moreno2Oscar E Simonson3Sergey Rodin4Nathalie Teller5Karl Tryggvason6Karin E Lundin7Leonard Girnita8Carl Inge Edvard Smith9Department of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, Huddinge, SwedenDepartment of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, Huddinge, SwedenDepartment of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, Huddinge, SwedenDepartment of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, Huddinge, SwedenDivision of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SwedenEurogentec S.A., Seraing, BelgiumDivision of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SwedenDepartment of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, Huddinge, SwedenDepartment of Oncology and Pathology, Cancer Center Karolinska, Karolinska Institutet and Karolinska University Hospital, Stockholm, SwedenDepartment of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, Huddinge, SwedenInducible systems for gene expression emerge as a new class of artificial vectors offering temporal and spatial exogenous control of gene expression. However, most inducible systems are less efficient in vivo and lack the target-organ specificity. In the present study, we have developed and optimized an oligonucleotide-based inducible system for the in vivo control of transgenes in the liver. We generated a set of simple, inducible plasmid-vectors based on the addition of four units of liver-specific miR-122 target sites to the 3′untranslated region of the gene of interest. Once the vector was delivered into hepatocytes this modification induced a dramatic reduction of gene expression that could be restored by the infusion of an antagomir for miR-122. The efficiency of the system was tested in vivo, and displayed low background and strong increase in gene expression upon induction. Moreover, gene expression was repeatedly induced even several months after the first induction showing no toxic effect in vivo. By combining tissue-specific control elements with antagomir treatment we generated, optimized and validated a robust inducible system that could be used successfully for in vivo experimental models requiring tight and cyclic control of gene expression.http://www.sciencedirect.com/science/article/pii/S2162253116303146cell type specificliver expressionlong-term gene expressionreporter system |
| spellingShingle | Iulian I Oprea Joana R Viola Pedro M D Moreno Oscar E Simonson Sergey Rodin Nathalie Teller Karl Tryggvason Karin E Lundin Leonard Girnita Carl Inge Edvard Smith Repeatable, Inducible Micro-RNA-Based Technology Tightly Controls Liver Transgene Expression Molecular Therapy: Nucleic Acids cell type specific liver expression long-term gene expression reporter system |
| title | Repeatable, Inducible Micro-RNA-Based Technology Tightly Controls Liver Transgene Expression |
| title_full | Repeatable, Inducible Micro-RNA-Based Technology Tightly Controls Liver Transgene Expression |
| title_fullStr | Repeatable, Inducible Micro-RNA-Based Technology Tightly Controls Liver Transgene Expression |
| title_full_unstemmed | Repeatable, Inducible Micro-RNA-Based Technology Tightly Controls Liver Transgene Expression |
| title_short | Repeatable, Inducible Micro-RNA-Based Technology Tightly Controls Liver Transgene Expression |
| title_sort | repeatable inducible micro rna based technology tightly controls liver transgene expression |
| topic | cell type specific liver expression long-term gene expression reporter system |
| url | http://www.sciencedirect.com/science/article/pii/S2162253116303146 |
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