Generation of reproductive transgenic pigs of a CRISPR-Cas9-based oncogene-inducible system by somatic cell nuclear transfer

Kiyoung Eun, Seon Ung Hwang, Mirae Kim, Junchul David Yoon, Eunhye Kim, Hyerin Choi, Gahye Kim, Hee Young Jeon, Jun Kyum Kim, Jung Yun Kim, Nayoung Hong, Min Gi Park, Junseok Jang, Hyeon Ju Jeong, Sung Jin Kim, Bong Woo Ko, Sang Chul Lee, Hyunggee Kim, Sang Hwan Hyun

Research output: Contribution to journalArticlepeer-review

Abstract

Alternative cancer models that are close to humans are required to create more valuable preclinical results during oncology studies. Here, a new onco-pig model via developing a CRISPR-Cas9-based Conditional Polycistronic gene expression Cassette (CRI-CPC) system to control the tumor inducing simian virus 40 large T antigen (SV40LT) and oncogenic HRASG12V. After conducting somatic cell nuclear transfer (SCNT), transgenic embryos were transplanted into surrogate mothers and five male piglets were born. Umbilical cord analysis confirmed that all piglets were transgenic. Two of them survived and they expressed a detectable green fluorescence. The test was made whether CRI-CPC models were naturally fertile and whether the CRI-CPC system was stably transferred to the offspring. By mating with a normal female pig, four offspring piglets were successfully produced. Among them, only three male piglets were transgenic. Finally, their applicability was tested as cancer models after transduction of Cas9 into fibroblasts from each CRI-CPC pig in vitro, resulting in cell acquisition of cancerous characteristics via the induction of oncogene expression. These results showed that our new CRISPR-Cas9-based onco-pig model was successfully developed.

Original languageEnglish
JournalBiotechnology Journal
DOIs
Publication statusAccepted/In press - 2022

Keywords

  • CRISPR-Cas9
  • pig
  • somatic cell nuclear transfer
  • transgenic cancer model

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Molecular Medicine

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