Targeted, homology-driven gene insertion in stem cells by ZFN-loaded ‘all-in-one’ lentiviral vectors
| Autor: | Jacob Giehm Mikkelsen, Yonglun Luo, Martin R. Jakobsen, Mads Valdemar Anderson, Yan Zhou, Yujia Cai, Shengting Li, Niels Uldbjerg, Anders Laustsen, Chenglong Sun |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
QH301-705.5 Genetic enhancement Science Genetic Vectors Biology General Biochemistry Genetics and Molecular Biology Viral vector protein transduction 03 medical and health sciences Humans Insertion Biology (General) Homologous Recombination Induced pluripotent stem cell Cells Cultured Genetics Deoxyribonucleases iPSC General Immunology and Microbiology Stem Cells General Neuroscience Lentivirus lentiviral vector Gene targeting Genetic Therapy General Medicine Zinc finger nuclease CD34+ cells Virus Cell biology Mutagenesis Insertional DNA Repair Enzymes Developmental Biology and Stem Cells 030104 developmental biology Genes and Chromosomes Gene Targeting zinc-finger nucleases Medicine Stem cell Research Advance Homologous recombination Human |
| Zdroj: | eLife, Vol 5 (2016) Cai, Y, Laustsen, A, Zhou, Y, Sun, C, Anderson, M V, Li, S, Uldbjerg, N, Luo, Y, Jakobsen, M R & Giehm Mikkelsen, J 2016, ' Targeted, homology-driven gene insertion in stem cells by ZFN-loaded 'all-in-one' lentiviral vectors ', eLife, vol. 5, e12213 . https://doi.org/10.7554/eLife.12213 eLife Cai, Y, Laustsen, A, Zhou, Y, Sun, C, Anderson, M V, Li, S, Uldbjerg, N, Luo, Y, Jakobsen, M R & Mikkelsen, J G 2016, ' Targeted, homology-driven gene insertion in stem cells by ZFN-loaded 'all-in-one' lentiviral vectors ', eLife, vol. 5 . https://doi.org/10.7554/eLife.12213 |
| DOI: | 10.7554/eLife.12213 |
| Popis: | Biased integration remains a key challenge for gene therapy based on lentiviral vector technologies. Engineering of next-generation lentiviral vectors targeting safe genomic harbors for insertion is therefore of high relevance. In a previous paper (Cai et al., 2014a), we showed the use of integrase-defective lentiviral vectors (IDLVs) as carriers of complete gene repair kits consisting of zinc-finger nuclease (ZFN) proteins and repair sequences, allowing gene correction by homologous recombination (HR). Here, we follow this strategy to engineer ZFN-loaded IDLVs that insert transgenes by a homology-driven mechanism into safe loci. This insertion mechanism is driven by time-restricted exposure of treated cells to ZFNs. We show targeted gene integration in human stem cells, including CD34+ hematopoietic progenitors and induced pluripotent stem cells (iPSCs). Notably, targeted insertions are identified in 89% of transduced iPSCs. Our findings demonstrate the applicability of nuclease-loaded ‘all-in-one’ IDLVs for site-directed gene insertion in stem cell-based gene therapies. |
| Databáze: | OpenAIRE |
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