Efficient generation of transgenic cattle using the DNA transposon and their analysis by next-generation sequencing.
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.
Department of Theriogenology, College of Veterinary Medicine and the Research Institute of Veterinary Science, Seoul National University, 08826, Republic of Korea.
Soo-Young
SY
Yum
Embryo Research Center, Seoul Milk Coop, Gyeonggi-do, 12528, Republic of Korea.
Song-Jeon
SJ
Lee
TheragenEtex BiO Institute, Advanced Institutes of Convergence Technology, Kwanggyo Technovalley, Suwon, 16229, Republic of Korea.
Hyun-Min
HM
Kim
Department of Theriogenology, College of Veterinary Medicine and the Research Institute of Veterinary Science, Seoul National University, 08826, Republic of Korea.
Woo-Jae
WJ
Choi
Department of Theriogenology, College of Veterinary Medicine and the Research Institute of Veterinary Science, Seoul National University, 08826, Republic of Korea.
Ji-Hyun
JH
Park
Embryo Research Center, Seoul Milk Coop, Gyeonggi-do, 12528, Republic of Korea.
Won-Wu
WW
Lee
Embryo Research Center, Seoul Milk Coop, Gyeonggi-do, 12528, Republic of Korea.
Hee-Soo
HS
Kim
Embryo Research Center, Seoul Milk Coop, Gyeonggi-do, 12528, Republic of Korea.
Hyeong-Jong
HJ
Kim
Embryo Research Center, Seoul Milk Coop, Gyeonggi-do, 12528, Republic of Korea.
Seong-Hun
SH
Bae
Embryo Research Center, Seoul Milk Coop, Gyeonggi-do, 12528, Republic of Korea.
Je-Hyeong
JH
Lee
Embryo Research Center, Seoul Milk Coop, Gyeonggi-do, 12528, Republic of Korea.
Joo-Yeong
JY
Moon
Department of Theriogenology, College of Veterinary Medicine and the Research Institute of Veterinary Science, Seoul National University, 08826, Republic of Korea.
Ji-Hyun
JH
Lee
Department of Theriogenology, College of Veterinary Medicine and the Research Institute of Veterinary Science, Seoul National University, 08826, Republic of Korea.|Department of Chemistry, College of Natural Science, Seoul National University, 08826, Republic of Korea.
Choong-Il
CI
Lee
TheragenEtex BiO Institute, Advanced Institutes of Convergence Technology, Kwanggyo Technovalley, Suwon, 16229, Republic of Korea.
Bong-Jun
BJ
Son
TheragenEtex BiO Institute, Advanced Institutes of Convergence Technology, Kwanggyo Technovalley, Suwon, 16229, Republic of Korea.
Sang-Hoon
SH
Song
TheragenEtex BiO Institute, Advanced Institutes of Convergence Technology, Kwanggyo Technovalley, Suwon, 16229, Republic of Korea.
Su-Min
SM
Ji
TheragenEtex BiO Institute, Advanced Institutes of Convergence Technology, Kwanggyo Technovalley, Suwon, 16229, Republic of Korea.
Seong-Jin
SJ
Kim
Department of Theriogenology, College of Veterinary Medicine and the Research Institute of Veterinary Science, Seoul National University, 08826, Republic of Korea.|Emergence Center for Food-Medicine Personalized Therapy System, Advanced Institutes of Convergence Technology, Seoul National University, Gyeonggi-do, 16229, Republic of Korea.
Goo
G
Jang
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics* / Q000254:growth & development; D001755:Blastocyst / Q000378:metabolism; D002417:Cattle; D004251:DNA Transposable Elements / Q000235:genetics*; D005260:Female; D042822:Genomic Instability; D059014:High-Throughput Nucleotide Sequencing; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism; D019076:Transgenes / Q000235:genetics*; D019895:Transposases / Q000235:genetics
D004251:DNA Transposable Elements; D019895:Transposases