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Reactivation of Transgene Expression by Alleviating CpG Methylation of the Rous sarcoma virus Promoter in Transgenic Quail Cells

Cited 4 time in Web of Science Cited 4 time in Scopus
Authors
Jang, Hyun-Jun; Choi, Jin Won; Kim, Young Min; Shin, Sang Su; Lee, Kichoon; Han, Jae Yong
Issue Date
2011
Publisher
Humana Press
Citation
Molecular Biotechnology, vol.49 no.3, pp. 222-228
Keywords
Transgenic quailRSV promoterTransgene expressionCpG methylation5-aza-20-deoxycytidine
Abstract
In this study, we investigated the relative expression of the Rous sarcoma virus (RSV) promoterdriven expression of enhanced green fluorescent protein (EGFP) in fibroblasts of transgenic quails. We analyzed the direct influence of CpG methylation of the RSV promoter on the transcriptional activity of delivered transgenes. Embryonic fibroblasts collected from homozygous transgenic quail (TQ2) were treated with 50 lM of DNA methyltransferase inhibitor followed by 5-aza-20-deoxycytidine (5-azadC) for 48 h, and changes in expression were then analyzed by flow cytometry. The results show a significant increase of EGFP expression in TQ2 embryonic fibroblasts (QEFs) (2.64% to 79.84%). Subsequent methylation- specific amplification revealed that 5-azadC significantly reduced the CpG methylation status in the RSV promoters of the QEFs (86.42 to 48.41%); even after 5-azadC was withdrawn, CpG methylation remained decreased in expanded culture (16.28%). Further analysis showed that potential transcription factor binding sites existed in the CpG methylation site of the RSV promoter. These results may provide the basis for understanding the epigenetic mechanism responsible for transgenic animal production and genetic preservation.
ISSN
1073-6085
Language
English
URI
https://hdl.handle.net/10371/100315
DOI
https://doi.org/10.1007/s12033-011-9393-7
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College of Agriculture and Life Sciences (농업생명과학대학)Dept. of Food and Animal Biotechnology (식품·동물생명공학부)Journal Papers (저널논문_식품·동물생명공학부)
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