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KMID : 0894520130170040379
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Development & Reproduction
2013 Volume.17 No. 4 p.379 ~ p.387
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Enhancement of Transgene Expression by HDAC Inhibitors in Mouse Embryonic Stem Cells
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Kim Young-Eun
Park Jeong-A
Park Sang-Kyu
Kang Ho-Bum
Kwon Hyung-Joo
Lee Young-Hee
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Abstract
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Embryonic stem (ES) cells can self-renew and differentiate to various cells depending on the culture condition. Although ES cells are a good model for cell type specification and can be useful for application in clinics in the future, studies on ES cells have many experimental restraints including low transfection efficiency and transgene expression. Here, we observed that transgene expression after transfection was enhanced by treatment with histone deacetylse (HDAC) inhibitors such as trichostatin A, sodium butyrate, and valproic acid. Transfection was performed using conventional transfection reagents with a retroviral vector encoding GFP under the control of CMV promoter as a reporter. Treatment of ES cells with HDAC inhibitors after transfection increased population of GFP positive cells up to 180% compared with untreated control. ES cells showed normal expression of stem cell markers after treatment with HDAC inhibitors. Transgene expression was further enhanced by modifying transfection procedure. GFP positive cells selected after transfection were proved to have the stem cell properties. Our improved protocol for enhanced gene delivery and expression in mouse ES cells without hampering ES cell properties will be useful for study and application of ES cells.
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KEYWORD
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Mouse embryonic stem cell, HDAC inhibitor, Transfection, Enhancement, Optimization
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