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Improved efficiencies in the generation of multigene-modified pigs by recloning and using sows as the recipient

Published online by Cambridge University Press:  28 June 2021

Jongki Cho Open the ORCID record for Jongki Cho [Opens in a new window] ,
Ghangyong Kim ,
Ahmad Yar Qamar ,
Xun Fang ,
Pantu Kumar Roy ,
Bereket Molla Tanga ,
Seonggyu Bang ,
Jong Keun Kim ,
Cesare Galli  and
Andrea Perota
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Jongki Cho
Affiliation:
College of Veterinary Medicine, Chungnam National University, Daejeon34134, Republic of Korea
Ghangyong Kim
Affiliation:
College of Veterinary Medicine, Chungnam National University, Daejeon34134, Republic of Korea
Ahmad Yar Qamar
Affiliation:
College of Veterinary Medicine, Chungnam National University, Daejeon34134, Republic of Korea
Xun Fang
Affiliation:
College of Veterinary Medicine, Chungnam National University, Daejeon34134, Republic of Korea
Pantu Kumar Roy
Affiliation:
College of Veterinary Medicine, Chungnam National University, Daejeon34134, Republic of Korea
Bereket Molla Tanga
Affiliation:
College of Veterinary Medicine, Chungnam National University, Daejeon34134, Republic of Korea
Seonggyu Bang
Affiliation:
College of Veterinary Medicine, Chungnam National University, Daejeon34134, Republic of Korea
Jong Keun Kim
Affiliation:
Department of Animal Resource and Science, Dankook University, Cheonan-si, Chungnam 31116, Republic of Korea
Cesare Galli
Affiliation:
Avantea, Laboratory of Reproductive Technologies, Via Porcellasco 7/f, Cremona26100, Italy
Andrea Perota
Affiliation:
Avantea, Laboratory of Reproductive Technologies, Via Porcellasco 7/f, Cremona26100, Italy
Young Tae Kim
Affiliation:
Department of Thoracic and Cardiovascular Surgery, Seoul National University Hospital, Seoul National University College of Medicine, Seoul03080, Republic of Korea
Jeong-Hwan Che
Affiliation:
Biomedical Center for Animal Resource and Development, Seoul National University College of Medicine, Seoul03080, Republic of Korea
Chung-Gyu Park*
Affiliation:
Department of Microbiology and Immunology; Xenotransplantation Research Center, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea
*
Author for correspondence: Chung-Gyu Park. Department of Microbiology and Immunology; Xenotransplantation Research Center, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea. Email: chgpark@snu.ac.kr
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Summary

This study was performed to improve production efficiency at the level of recipient pig and donor nuclei of transgenic cloned pigs used for xenotransplantation. To generate transgenic pigs, human endothelial protein C receptor (hEPCR) and human thrombomodulin (hTM) genes were introduced using the F2A expression vector into GalT–/–/hCD55+ porcine neonatal ear fibroblasts used as donor cells and cloned embryos were transferred to the sows and gilts. Cloned fetal kidney cells were also used as donor cells for recloning to increase production efficiency. Pregnancy and parturition rates after embryo transfer and preimplantation developmental competence were compared between cloned embryos derived from adult and fetal cells. Significantly higher parturition rates were shown in the group of sows (50.0 vs. 4.1%), natural oestrus (20.8 vs. 0%), and ovulated ovary (16.7 vs. 5.6%) compared with gilt, induced and non-ovulated, respectively (P < 0.05). When using gilts as recipients, final parturitions occurred in only the fetal cell groups and significantly higher blastocyst rates (15.1% vs. 21.3%) were seen (P < 0.05). Additionally, gene expression levels related to pluripotency were significantly higher in the fetal cell group (P < 0.05). In conclusion, sows can be recommended as recipients due to their higher efficiency in the generation of transgenic cloned pigs and cloned fetal cells also can be recommended as donor cells through correct nuclear reprogramming.

Keywords

PorcineRecloningRecipientReprogrammingTransgenic
Type
Research Article
Information
Zygote , Volume 30 , Issue 1 , February 2022 , pp. 103 - 110
Copyright
© The Author(s), 2021. Published by Cambridge University Press

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