Novel Non-integrating DNA Nano-S/MAR Vectors Restore Gene Function in Isogenic Patient-Derived Pancreatic Tumor Models
| Autor: | Williams James A, Martin R. Sprick, Alice De Roia, Vanessa Vogel, Richard Harbottle, Rienk Offringa, Edward W. Green, Corinna Klein, Matthias Bozza, Elisa Espinet |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Tumor suppressor gene lcsh:QH426-470 Transgene Cell Biology antibiotic-free isogenic cells Article S/MAR Transcriptome Insertional mutagenesis non-integrating 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Genetics medicine lcsh:QH573-671 Molecular Biology Mitosis Gene lcsh:Cytology tumor models gene supplementation nano-DNA vector Cell biology lcsh:Genetics 030104 developmental biology medicine.anatomical_structure chemistry 030220 oncology & carcinogenesis Molecular Medicine DNA |
| Zdroj: | Molecular Therapy. Methods & Clinical Development Molecular Therapy: Methods & Clinical Development, Vol 17, Iss, Pp 957-968 (2020) |
| ISSN: | 2329-0501 |
| Popis: | We describe herein non-integrating minimally sized nano-S/MAR DNA vectors, which can be used to genetically modify dividing cells in place of integrating vectors. They represent a unique genetic tool, which avoids vector-mediated damage. Previous work has shown that DNA vectors comprising a mammalian S/MAR element can provide persistent mitotic stability over hundreds of cell divisions, resisting epigenetic silencing and thereby allowing sustained transgene expression. The composition of the original S/MAR vectors does present some inherent limitations that can provoke cellular toxicity. Herein, we present a new system, the nano-S/MAR, which drives higher transgene expression and has improved efficiency of establishment, due to the minimal impact on cellular processes and perturbation of the endogenous transcriptome. We show that these features enable the hitherto challenging genetic modification of patient-derived cells to stably restore the tumor suppressor gene SMAD4 to a patient-derived SMAD4 knockout pancreatic cancer line. Nano-S/MAR modification does not alter the molecular or phenotypic integrity of the patient-derived cells in cell culture and xenograft mouse models. In conclusion, we show that these DNA vectors can be used to persistently modify a range of cells, providing sustained transgene expression while avoiding the risks of insertional mutagenesis and other vector-mediated toxicity. Graphical Abstract Bozza and et al. describe minimally sized DNA vectors that can be used to generate genetically modified isogenic human cells that produce persistent transgene expression while avoiding insertional mutagenesis and vector-mediated toxicity. They show the restoration of the tumor suppressor gene SMAD4 into patient-derived pancreatic cells reversing the cellular and molecular phenotype. |
| Databáze: | OpenAIRE |
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