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Transfer of synthetic human chromosome into human induced pluripotent stem cells for biomedical applications

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Item Type:Article
Title:Transfer of synthetic human chromosome into human induced pluripotent stem cells for biomedical applications
Creators Name:Sinenko, S.A., Skvortsova, E.V., Liskovykh, M.A., Ponomartsev, S.V., Kuzmin, A.A., Khudiakov, A.A., Malashicheva, A.B., Alenina, N., Larionov, V., Kouprina, N. and Tomilin, A.N.
Abstract:Alphoid(tetO)-type human artificial chromosome (HAC) has been recently synthetized as a novel class of gene delivery vectors for induced pluripotent stem cell (iPSC)-based tissue replacement therapeutic approach. This HAC vector was designed to deliver copies of genes into patients with genetic diseases caused by the loss of a particular gene function. The alphoid(tetO)-HAC vector has been successfully transferred into murine embryonic stem cells (ESCs) and maintained stably as an independent chromosome during the proliferation and differentiation of these cells. Human ESCs and iPSCs have significant differences in culturing conditions and pluripotency state in comparison with the murine naïve-type ESCs and iPSCs. To date, transferring alphoid(tetO)-HAC vector into human iPSCs (hiPSCs) remains a challenging task. In this study, we performed the microcell-mediated chromosome transfer (MMCT) of alphoid(tetO)-HAC expressing the green fluorescent protein into newly generated hiPSCs. We used a recently modified MMCT method that employs an envelope protein of amphotropic murine leukemia virus as a targeting cell fusion agent. Our data provide evidence that a totally artificial vector, alphoid(tetO)-HAC, can be transferred and maintained in human iPSCs as an independent autonomous chromosome without affecting pluripotent properties of the cells. These data also open new perspectives for implementing alphoid(tetO)-HAC as a gene therapy tool in future biomedical applications.
Keywords:Human Artificial Chromosome (HAC), Alphoid(tetO)-HAC, Induced Pluripotent Stem Cells (iPSCs), Microcell-Mediated Chromosome Transfer (MMCT), Cell Reprogramming
Source:Cells
ISSN:2073-4409
Publisher:MDPI
Volume:7
Number:12
Page Range:261
Date:8 December 2018
Official Publication:https://doi.org/10.3390/cells7120261
PubMed:View item in PubMed

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