Development of a Positive-readout Mouse Model of siRNA Pharmacodynamics
Development of RNAi-based therapeutics has the potential to revolutionize treatment options for a range of human diseases. However, as with gene therapy, a major barrier to progress is the lack of methods to achieve and measure efficient delivery for systemic administration. We have developed a posi...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2013-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253116301949 |
| _version_ | 1818416422580125696 |
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| author | Mark Stevenson Robert Carlisle Ben Davies Chris Preece Michelle Hammett Wei-li Liu Kerry David Fisher Amy Ryan Heidi Scrable Leonard William Seymour |
| author_facet | Mark Stevenson Robert Carlisle Ben Davies Chris Preece Michelle Hammett Wei-li Liu Kerry David Fisher Amy Ryan Heidi Scrable Leonard William Seymour |
| author_sort | Mark Stevenson |
| collection | DOAJ |
| description | Development of RNAi-based therapeutics has the potential to revolutionize treatment options for a range of human diseases. However, as with gene therapy, a major barrier to progress is the lack of methods to achieve and measure efficient delivery for systemic administration. We have developed a positive-readout pharmacodynamic transgenic reporter mouse model allowing noninvasive real-time assessment of siRNA activity. The model combines a luciferase reporter gene under the control of regulatory elements from the lac operon of Escherichia coli. Introduction of siRNA targeting lac repressor results in increased luciferase expression in cells where siRNA is biologically active. Five founder luciferase-expressing and three founder Lac-expressing lines were generated and characterized. Mating of ubiquitously expressing luciferase and lac lines generated progeny in which luciferase expression was significantly reduced compared with the parental line. Administration of isopropyl β-D-1-thiogalactopyranoside either in drinking water or given intraperitoneally increased luciferase expression in eight of the mice examined, which fell rapidly when withdrawn. Intraperitoneal administration of siRNA targeting lac in combination with Lipofectamine 2000 resulted in increased luciferase expression in the liver while control nontargeting siRNA had no effect. We believe a sensitive positive readout pharmacodynamics reporter model will be of use to the research community in RNAi-based vector development. |
| first_indexed | 2024-12-14T11:50:38Z |
| format | Article |
| id | doaj.art-663304a2de414c409ddb14be890b7809 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-14T11:50:38Z |
| publishDate | 2013-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-663304a2de414c409ddb14be890b78092022-12-21T23:02:20ZengElsevierMolecular Therapy: Nucleic Acids2162-25312013-01-012C10.1038/mtna.2013.63Development of a Positive-readout Mouse Model of siRNA PharmacodynamicsMark Stevenson0Robert Carlisle1Ben Davies2Chris Preece3Michelle Hammett4Wei-li Liu5Kerry David Fisher6Amy Ryan7Heidi Scrable8Leonard William Seymour9Academic Endocrine Unit, OCDEM, University of Oxford, Oxford, UKInstitute of Biomedical Engineering, University of Oxford, Oxford, UKTransgenic Core, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UKTransgenic Core, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UKTransgenic Core, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, UKDepartment of Oncology, University of Oxford, Oxford, UKDepartment of Oncology, University of Oxford, Oxford, UKClinton Community College, Plattsburgh, New York, USADepartment of Biochemistry/Molecular Biology, College of Medicine, Mayo Clinic, Rochester, Minnesota, USADepartment of Oncology, University of Oxford, Oxford, UKDevelopment of RNAi-based therapeutics has the potential to revolutionize treatment options for a range of human diseases. However, as with gene therapy, a major barrier to progress is the lack of methods to achieve and measure efficient delivery for systemic administration. We have developed a positive-readout pharmacodynamic transgenic reporter mouse model allowing noninvasive real-time assessment of siRNA activity. The model combines a luciferase reporter gene under the control of regulatory elements from the lac operon of Escherichia coli. Introduction of siRNA targeting lac repressor results in increased luciferase expression in cells where siRNA is biologically active. Five founder luciferase-expressing and three founder Lac-expressing lines were generated and characterized. Mating of ubiquitously expressing luciferase and lac lines generated progeny in which luciferase expression was significantly reduced compared with the parental line. Administration of isopropyl β-D-1-thiogalactopyranoside either in drinking water or given intraperitoneally increased luciferase expression in eight of the mice examined, which fell rapidly when withdrawn. Intraperitoneal administration of siRNA targeting lac in combination with Lipofectamine 2000 resulted in increased luciferase expression in the liver while control nontargeting siRNA had no effect. We believe a sensitive positive readout pharmacodynamics reporter model will be of use to the research community in RNAi-based vector development.http://www.sciencedirect.com/science/article/pii/S2162253116301949Lac operonLac repressortransgenic reporter mouse |
| spellingShingle | Mark Stevenson Robert Carlisle Ben Davies Chris Preece Michelle Hammett Wei-li Liu Kerry David Fisher Amy Ryan Heidi Scrable Leonard William Seymour Development of a Positive-readout Mouse Model of siRNA Pharmacodynamics Molecular Therapy: Nucleic Acids Lac operon Lac repressor transgenic reporter mouse |
| title | Development of a Positive-readout Mouse Model of siRNA Pharmacodynamics |
| title_full | Development of a Positive-readout Mouse Model of siRNA Pharmacodynamics |
| title_fullStr | Development of a Positive-readout Mouse Model of siRNA Pharmacodynamics |
| title_full_unstemmed | Development of a Positive-readout Mouse Model of siRNA Pharmacodynamics |
| title_short | Development of a Positive-readout Mouse Model of siRNA Pharmacodynamics |
| title_sort | development of a positive readout mouse model of sirna pharmacodynamics |
| topic | Lac operon Lac repressor transgenic reporter mouse |
| url | http://www.sciencedirect.com/science/article/pii/S2162253116301949 |
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