The β-Globin Locus Control Region in Combination With the EF1α Short Promoter Allows Enhanced Lentiviral Vector-mediated Erythroid Gene Expression With Conserved Multilineage Activity
| Autor: | Aiste Monkeviciute, ME Alonso-Ferrero, Simone Scholz, Reiner Schulz, H. Bobby Gaspar, Marlene Carmo, Lin Zhang, Heba Saadeh, Manfred G. Schmidt, Yasuhiro Takeuchi, Michael Antoniou, Adrian J. Thrasher, Sean Knight, Lynette D. Fairbanks, Claudia Montiel-Equihua, Michael P. Blundell, Mary Collins |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Adenosine Deaminase
Transgene Genetic enhancement Genetic Vectors Green Fluorescent Proteins beta-Globins Biology Viral vector Cell Line 03 medical and health sciences Jurkat Cells Mice 0302 clinical medicine Peptide Elongation Factor 1 Gene expression Drug Discovery Transcriptional regulation Genetics Animals Humans Promoter Regions Genetic Molecular Biology Erythroid Precursor Cells Locus control region 030304 developmental biology Regulation of gene expression Pharmacology 0303 health sciences Lentivirus Genetic Therapy U937 Cells Hematopoietic Stem Cells Locus Control Region Peptide Elongation Factors Molecular biology Up-Regulation Lysosomal Storage Diseases Mice Inbred C57BL HEK293 Cells Gene Expression Regulation Molecular Medicine Original Article 030217 neurology & neurosurgery |
| Zdroj: | Molecular Therapy; Vol 20 |
| ISSN: | 1525-0016 |
| DOI: | 10.1038/mt.2012.50 |
| Popis: | Some gene therapy strategies are compromised by the levels of gene expression required for therapeutic benefit, and also by the breadth of cell types that require correction. We designed a lentiviral vector system in which a transgene is under the transcriptional control of the short form of constitutively acting elongation factor 1α promoter (EFS) combined with essential elements of the locus control region of the β-globin gene (β-LCR). We show that the β-LCR can upregulate EFS activity specifically in erythroid cells but does not alter EFS activity in myeloid or lymphoid cells. Experiments using the green fluorescent protein (GFP) reporter or the human adenosine deaminase (ADA) gene demonstrate 3–7 times upregulation in vitro but >20 times erythroid-specific upregulation in vivo, the effects of which were sustained for 1 year. The addition of the β-LCR did not alter the mutagenic potential of the vector in in vitro mutagenesis (IM) assays although microarray analysis showed that the β-LCR upregulates ~9% of neighboring genes. This vector design therefore combines the benefits of multilineage gene expression with high-level erythroid expression, and has considerable potential for correction of multisystem diseases including certain lysosomal storage diseases through a hematopoietic stem cell (HSC) gene therapy approach. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|