title: Enhancement of Transgene Expression by HDAC Inhibitors in Mouse Embryonic Stem Cells.
abstract: 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.
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Republic of Korea ; Biotechnology Research Institute, Chungbuk National University, Cheongju 361-763, Republic of Korea.
authors.forename: Young-Eun
authors.initials: YE
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Republic of Korea.
authors.forename: Jeong-A
authors.initials: JA
authors.lastname: Park
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Republic of Korea.
authors.forename: Sang-Kyu
authors.initials: SK
authors.lastname: Park
authors.affiliation: Biotechnology Research Institute, Chungbuk National University, Cheongju 361-763, Republic of Korea.
authors.forename: Ho-Bum
authors.initials: HB
authors.lastname: Kang
authors.affiliation: Center for Medical Science Research, Republic of Korea ; Department of Microbiology, College of Medicine, Hallym University, Chuncheon 200-702, Republic of Korea.
authors.forename: Hyung-Joo
authors.initials: HJ
authors.lastname: Kwon
authors.affiliation: Department of Biochemistry, College of Natural Sciences, Republic of Korea ; Biotechnology Research Institute, Chungbuk National University, Cheongju 361-763, Republic of Korea.
authors.forename: Younghee
authors.initials: Y
authors.lastname: Lee
keywords: Enhancement; HDAC inhibitor; Mouse embryonic stem cell; Optimization.; Transfection