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Issue 46, 2014
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Enhanced non-viral gene delivery to human embryonic stem cells via small molecule-mediated transient alteration of the cell structure

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Abstract

Non-viral gene delivery into human embryonic stem cells (hESCs) is an important tool for controlling cell fate. However, the delivery efficiency remains low due in part to the tight colony structure of the cells which prevents effective exposure towards delivery vectors. We herein report a novel approach to enhance non-viral gene delivery to hESCs by transiently altering the cell and colony structure. (R)-(+)-trans-4-(1-aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide (Y-27632), a small molecule that inhibits the rho-associated protein kinase pathway, is utilized to induce transient colony spreading which leads to an increased transfection efficiency by 1.5 to 2 folds in a spectrum of non-viral transfection reagents including Lipofectamine 2000 and Fugene HD. After removal of Y-27632 post-transfection, cells can revert back to their normal state and do not show alteration of pluripotency. This approach provides a simple, effective tool to enhance non-viral gene delivery into adherent hESCs for genetic manipulation.

Graphical abstract: Enhanced non-viral gene delivery to human embryonic stem cells via small molecule-mediated transient alteration of the cell structure

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Publication details

The article was received on 09 May 2014, accepted on 13 Sep 2014 and first published on 15 Sep 2014


Article type: Communication
DOI: 10.1039/C4TB00750F
Author version available: Download Author version (PDF)
Citation: J. Mater. Chem. B, 2014,2, 8098-8105
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    Enhanced non-viral gene delivery to human embryonic stem cells via small molecule-mediated transient alteration of the cell structure

    J. Yen, L. Yin and J. Cheng, J. Mater. Chem. B, 2014, 2, 8098
    DOI: 10.1039/C4TB00750F

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