Expression of 2A peptide mediated tri-fluorescent protein genes were regulated by epigenetics in transgenic sheep

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• 作者：Yongzhi Tian^a ; ^b ; Wenrong Li^b ; ^c ; ^d ; Liqin Wang^b ; ^c ; ^d ; Chenxi Liu^b ; ^c ; ^d ; Jiapeng Lin^b ; ^c ; ^d ; Xuemei Zhang^b ; ^c ; ^d ; Ning Zhang^b ; ^c ; ^d ; Sangang He^b ; ^c ; ^d ; Juncheng Huang^b ; ^c ; ^d ; Bin Jia^a ; Mingjun Liu^b ; ^c ; ^d ; ^{mingjun_l@sina.com}
• 关键词：2A peptide ; Transgenic sheep ; DNA methylation ; Lentiviral vector
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：2013
• 出版时间：10 May, 2013
• 年：2013
• 卷：434
• 期：3
• 页码：681-687
• 全文大小：1681 K

文摘

A number of gene therapy applications and basic research would benefit from vectors expressing multiple genes. In this study, we constructed 2A peptide based tricistronic lentiviral vector and generated transgenic lambs by injecting lentivirus carrying the tricistronic vector into perivitelline space of zygotes. Of 7 lambs born, 2 lambs (#6 and #7) carried the transgene. However, no fluorescent proteins were identified in transgenic sheep. To investigate why the transgene was silenced in transgenic sheep, we analyzed the methylation status of transgene. The methylation level of CMV promoter was 76.25 % in #6, and 64.7 % in #7. In the coding region of three fluorescent protein genes, methylation levels were extremely high, with the average level of 98.3 % in #6 and 98.4 % in #7 respectively. Furthermore, the ratio of GFP⁺ cells were increased significantly when the fibroblasts derived from the transgenic sheep were treated with 5-azaC and/ or TSA. Our results showed that 2A peptide based tricistronic construct was subjected to hypermethylation in transgenic sheep. Moreover, the silencing could be relieved by treating with methytransferase inhibitor and/or deacetylase inhibitor.