Efficient generation of transgenic cattle using the DNA transposon and their analysis by next-generation sequencing

利用 DNA 转座子高效生成转基因牛并通过下一代测序进行分析

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作者:Soo-Young Yum, Song-Jeon Lee, Hyun-Min Kim, Woo-Jae Choi, Ji-Hyun Park, Won-Wu Lee, Hee-Soo Kim, Hyeong-Jong Kim, Seong-Hun Bae, Je-Hyeong Lee, Joo-Yeong Moon, Ji-Hyun Lee, Choong-Il Lee, Bong-Jun Son, Sang-Hoon Song, Su-Min Ji, Seong-Jin Kim, Goo Jang

Abstract

Here, we efficiently generated transgenic cattle using two transposon systems (Sleeping Beauty and Piggybac) and their genomes were analyzed by next-generation sequencing (NGS). Blastocysts derived from microinjection of DNA transposons were selected and transferred into recipient cows. Nine transgenic cattle have been generated and grown-up to date without any health issues except two. Some of them expressed strong fluorescence and the transgene in the oocytes from a superovulating one were detected by PCR and sequencing. To investigate genomic variants by the transgene transposition, whole genomic DNA were analyzed by NGS. We found that preferred transposable integration (TA or TTAA) was identified in their genome. Even though multi-copies (i.e. fifteen) were confirmed, there was no significant difference in genome instabilities. In conclusion, we demonstrated that transgenic cattle using the DNA transposon system could be efficiently generated, and all those animals could be a valuable resource for agriculture and veterinary science.

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