Persistent Integration of Reprogramming Factors Impairs the In Vitro Cardiogenic Potential of Induced Pluripotent Stem Cells

Mittal, Nikhil; Domian, Ibrahim J.

doi:10.1161/circgenetics.114.000825

Cited by 1 publication

(1 citation statement)

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“…Nanovectors express their transgenes at higher levels than traditional plasmid vectors [33,34]. As the reprogramming process requires a strong but transient expression of ectopic reprogramming factors [38][39][40], we reasoned that a nanovector may give a sufficient expression of reprogramming factors to reprogram cells and improve the safety of these vectors by removing the capacity for vector retention. Vectors were confirmed to be functional by individual transfection into HEK293T cells and Western blotting (Figure 1D,E).…”

Section: Generation Of a Novel Nonviral Reprogramming Platformmentioning

confidence: 99%

A Simple Nonviral Method to Generate Human Induced Pluripotent Stem Cells Using SMAR DNA Vectors

Hartley,

Burger,

Wincek

et al. 2024

Genes

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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.

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