Journal Article

DKFZ-2024-01113

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png A Simple Nonviral Method to Generate Human Induced Pluripotent Stem Cells Using SMAR DNA Vectors.

Hartley, A. (First author)DKFZ* ; Burger, L.DKFZ* ; Wincek, C.DKFZ* ; Dons, L. ; Li, T. ; Grewenig, A.DKFZ* ; Taşgın, T.DKFZ* ; Urban, M.DKFZ* ; Roig-Merino, A.DKFZ* ; Ghazvini, M. ; Harbottle, R. (Last author)DKFZ*

2024
MDPI Basel

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Please use a persistent id in citations: doi:10.3390/genes15050575

Abstract: 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.

Keyword(s): Induced Pluripotent Stem Cells: cytology (MeSH) ; Induced Pluripotent Stem Cells: metabolism (MeSH) ; Humans (MeSH) ; Genetic Vectors: genetics (MeSH) ; Cellular Reprogramming: genetics (MeSH) ; Fibroblasts: cytology (MeSH) ; Fibroblasts: metabolism (MeSH) ; Plasmids: genetics (MeSH) ; Epstein-Barr Virus Nuclear Antigens: genetics (MeSH) ; Cells, Cultured (MeSH) ; Transfection: methods (MeSH) ; S/MAR ; SMAR DNA vector ; iPSC ; nonviral ; reprogramming ; stem cells ; Epstein-Barr Virus Nuclear Antigens ; EBV-encoded nuclear antigen 1

Note: #EA:D420#LA:D420#

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Contributing Institute(s):

1. DNA-Vektoren (D420)

Research Program(s):

1. 314 - Immunologie und Krebs (POF4-314) (POF4-314)

Appears in the scientific report 2024

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