Efficient Non-viral Gene Delivery into Human Hematopoietic Stem Cells by Minicircle Sleeping Beauty Transposon Vectors
| العنوان: | Efficient Non-viral Gene Delivery into Human Hematopoietic Stem Cells by Minicircle Sleeping Beauty Transposon Vectors |
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| المؤلفون: | Carsten Rudolph, Johannes Geiger, Juan A. Bueren, Manish K. Aneja, Wolfgang Walther, Marta Holstein, Fulvio Mavilio, Juan Carlos Ordóñez Flores, Marco Schmeer, Valentina Poletti, Zsuzsanna Izsvák, Csaba Miskey, Dennis Kobelt, Martin Schleef, Halvard-Björn Bönig, Guillermo Guenechea, Esther Grueso, Elena Almarza, Cristina Mesa-Núñez, Zoltán Ivics |
| المساهمون: | Pierantoni-Morgagni Hospital, Partenaires INRAE, Approches génétiques intégrées et nouvelles thérapies pour les maladies rares (INTEGRARE), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Généthon, Inst. for Groundwater Management, Généthon, École Pratique des Hautes Études (EPHE) |
| المصدر: | Molecular Therapy Molecular Therapy, Cell Press, 2018, 26, pp.1137-1153. ⟨10.1016/j.ymthe.2018.01.012⟩ Molecular Therapy, 2018, 26, pp.1137-1153. ⟨10.1016/j.ymthe.2018.01.012⟩ |
| بيانات النشر: | HAL CCSD, 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0301 basic medicine, Transposable element, Cancer Research, Cell Survival, Genetic enhancement, [SDV]Life Sciences [q-bio], Genetic Vectors, chromosomal integration, gene therapy, gene vectors, hematopoietic stem cells, nonviral gene delivery, transposition, Animals, Flow Cytometry, Gene Expression, Hematopoietic Stem Cells, Humans, Mice, Mice, Knockout, Retroviridae, Transfection, Transgenes, DNA Transposable Elements, Gene Transfer Techniques, Biology, Gene delivery, Minicircle, 03 medical and health sciences, Drug Discovery, Genetics, ddc:610, Molecular Biology, Transposase, Pharmacology, Sleeping Beauty transposon system, 3. Good health, Cell biology, Transplantation, 030104 developmental biology, Cardiovascular and Metabolic Diseases, Molecular Medicine, Original Article |
| الوصف: | The Sleeping Beauty (SB) transposon system is a non-viral gene delivery platform that combines simplicity, inexpensive manufacture, and favorable safety features in the context of human applications. However, efficient correction of hematopoietic stem and progenitor cells (HSPCs) with non-viral vector systems, including SB, demands further refinement of gene delivery techniques. We set out to improve SB gene transfer into hard-to-transfect human CD34+ cells by vectorizing the SB system components in the form of minicircles that are devoid of plasmid backbone sequences and are, therefore, significantly reduced in size. As compared to conventional plasmids, delivery of the SB transposon system as minicircle DNA is ∼20 times more efficient, and it is associated with up to a 50% reduction in cellular toxicity in human CD34+ cells. Moreover, providing the SB transposase in the form of synthetic mRNA enabled us to further increase the efficacy and biosafety of stable gene delivery into hematopoietic progenitors ex vivo. Genome-wide insertion site profiling revealed a close-to-random distribution of SB transposon integrants, which is characteristically different from gammaretroviral and lentiviral integrations in HSPCs. Transplantation of gene-marked CD34+ cells in immunodeficient mice resulted in long-term engraftment and hematopoietic reconstitution, which was most efficient when the SB transposase was supplied as mRNA and nucleofected cells were maintained for 4–8 days in culture before transplantation. Collectively, implementation of minicircle and mRNA technologies allowed us to further refine the SB transposon system in the context of HSPC gene delivery to ultimately meet clinical demands of an efficient and safe non-viral gene therapy protocol. Graphical Abstract Ivics and collegues refined the Sleeping Beauty transposon system for gene transfer in human hematopoietic stem and progenitor cells by vectorizing the transposon components as minicircle DNA and synthetic mRNA. The advanced vector system enables efficient and safe non-viral engineering of hematopoietic cells that can be transplanted into immunodeficient mice. |
| وصف الملف: | application/octet-stream; application/pdf |
| اللغة: | English |
| تدمد: | 1525-0016 1525-0024 |
| DOI: | 10.1016/j.ymthe.2018.01.012⟩ |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a2efe0186b748c693b55bd9658567c5f https://hal.archives-ouvertes.fr/hal-02880799 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....a2efe0186b748c693b55bd9658567c5f |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15250016 15250024 |
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| DOI: | 10.1016/j.ymthe.2018.01.012⟩ |