TY  - JOUR
AU  - Hartley, Anna
AU  - Burger, Luisa
AU  - Wincek, Cornelia
AU  - Dons, Lieke
AU  - Li, Tracy
AU  - Grewenig, Annabel
AU  - Taşgın, Toros
AU  - Urban, Manuela
AU  - Roig-Merino, Alicia
AU  - Ghazvini, Mehrnaz
AU  - Harbottle, Richard
TI  - A Simple Nonviral Method to Generate Human Induced Pluripotent Stem Cells Using SMAR DNA Vectors.
JO  - Genes
VL  - 15
IS  - 5
SN  - 2073-4425
CY  - Basel
PB  - MDPI
M1  - DKFZ-2024-01113
SP  - 575
PY  - 2024
N1  - #EA:D420#LA:D420#
AB  - Induced pluripotent stem cells (iPSCs) are a powerful tool for biomedical research, but their production presents challenges and safety concerns. Yamanaka and Takahashi revolutionised the field by demonstrating that somatic cells could be reprogrammed into pluripotent cells by overexpressing four key factors for a sufficient time. iPSCs are typically generated using viruses or virus-based methods, which have drawbacks such as vector persistence, risk of insertional mutagenesis, and oncogenesis. The application of less harmful nonviral vectors is limited as conventional plasmids cannot deliver the levels or duration of the factors necessary from a single transfection. Hence, plasmids that are most often used for reprogramming employ the potentially oncogenic Epstein-Barr nuclear antigen 1 (EBNA-1) system to ensure adequate levels and persistence of expression. In this study, we explored the use of nonviral SMAR DNA vectors to reprogram human fibroblasts into iPSCs. We show for the first time that iPSCs can be generated using nonviral plasmids without the use of EBNA-1 and that these DNA vectors can provide sufficient expression to induce pluripotency. We describe an optimised reprogramming protocol using these vectors that can produce high-quality iPSCs with comparable pluripotency and cellular function to those generated with viruses or EBNA-1 vectors.
KW  - Induced Pluripotent Stem Cells: cytology
KW  - Induced Pluripotent Stem Cells: metabolism
KW  - Humans
KW  - Genetic Vectors: genetics
KW  - Cellular Reprogramming: genetics
KW  - Fibroblasts: cytology
KW  - Fibroblasts: metabolism
KW  - Plasmids: genetics
KW  - Epstein-Barr Virus Nuclear Antigens: genetics
KW  - Cells, Cultured
KW  - Transfection: methods
KW  - S/MAR (Other)
KW  - SMAR DNA vector (Other)
KW  - iPSC (Other)
KW  - nonviral (Other)
KW  - reprogramming (Other)
KW  - stem cells (Other)
KW  - Epstein-Barr Virus Nuclear Antigens (NLM Chemicals)
KW  - EBV-encoded nuclear antigen 1 (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:38790204
C2  - pmc:PMC11121542
DO  - DOI:10.3390/genes15050575
UR  - https://inrepo02.dkfz.de/record/290486
ER  -