Effect of trichostatin A on transfected donor cells and subsequent development of porcine cloned embryos.
Transgenes integrated into mammalian cells are silenced rapidly. This phenomenon correlates with repressed chromatin structure marked by histone hypoacetylation. This study investigated the effect of trichostatin A (TSA; a histone-deacetylase inhibitor) on EGFP expression in transfected cells and embryonic development after somatic cell nuclear transfer (SCNT). Porcine adult fibroblasts were transfected with a pEGFP-C1 vector. Then transfected cells, donor cells for SCNT, were pretreated with TSA, with the untreated cells being used as the control. Expression of EGFP in donor cells and reconstructed embryos was detected when exposed to blue light. Results showed that the percentage of EGFP-positive cells significantly increased when the transfected cells were treated with TSA and the increased expression of EGFP was sustained to at least the morula stage. In addition, the cytotoxic effect of TSA on the transfected cells was dose dependent. In conclusion, TSA can rescue the silenced EGFP gene. Even after transferring the TSA-treated cells to enucleated recipient oocytes, TSA retained the ability to rescue a silenced EGFP gene. In addition, TSA had an impact on cell proliferation.
Bo
Fu
F
College of Life Science, Northeast Agriculture University, Harbin 150030, PR China.
Di
Liu
L
Qing-chang
Fang
F
Liang
Ren
R
Hong
Ma
M
Liang
Wang
W
Zhen-hua
Guo
G
Zhong-qiu
Li
L
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology* / Q000187:drug effects*; D047108:Embryonic Development / Q000187:drug effects*; D005347:Fibroblasts / Q000187:drug effects; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006877:Hydroxamic Acids / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000187:drug effects*; D013552:Swine; D014162:Transfection; D019076:Transgenes
D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C012589:trichostatin A