Biochim Biophys Acta Mol Basis Dis. 2019 Feb 9. pii: S0925-4439(19)30043-2. doi: 10.1016/j.bbadis.2019.01.031. [Epub ahead of print]

Generation of induced pluripotent stem cells using elastin like polypeptides as a non-viral gene delivery system.

Lee CH¹, Ingrole RSJ¹, Gill HS².

Abstract

Induced pluripotent stem cells (iPSCs) have been generated from various somatic cells using different approaches; however, a major restriction of reprogramming methods is the use of viral vectors, which have the risk of causing genome-integration of viral DNA. Here, without a viral vector, we generated iPSCs from mouse fibroblasts using an elastin-like polypeptide (ELP)-based transfection method. Our findings support the possible use of ELPs for delivery of the reprogramming genes in to somatic cells for generation of iPSCs. Results of gel retardation assay demonstrated efficient complexation of ELPs with a plasmid containing the four Yamanaka stem cell factors, Oct-4, Klf4, c-myc, and Sox2. After transfection, the iPSCs showed embryonic stem cell-like characteristics, including expression of endogenous pluripotency genes, differentiation into three germ layer lineages, and formation of teratomas in vivo. Our results demonstrate that ELP-based gene delivery may provide a safe method for use in generation of virus-free and exogenous DNA-free iPSCs, which will be crucial for future applications in stem cell-based therapies.

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