Production of cloned genetically modified pigs by a new drug-free gene-modified cell isolation system

2014 Fiscal Year Final Research Report

• Project/Area Number: 24580411
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied animal science
• Research Institution: Kagoshima University
• Principal Investigator: SATO Masahiro 鹿児島大学, 医用ミニブタ・先端医療開発研究センター, 教授 (30287099)
• Co-Investigator(Kenkyū-buntansha): MIYOSHI Kazuchika 鹿児島大学農学部, 生物生産学科, 教授 (70363611)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: クローンブタ / 遺伝子導入 / ＫＯ / 遺伝子改変

Outline of Final Research Achievements

Our purpose of this study is to produce genetically modified piglets that are used for xenotransplantation. α-GalT is known to synthesize α-Gal epitope (α-Gal), which is a causative for hyperacute rejection of pig grafts. We first disrupted the α-GalT gene in porcine cells using a recently developed technology that allows efficient ablation of a target gene, and finally obtained a cell line called CRISPR-2-1 (2-1). DAF and TM are also known as key factors suppressing graft rejection. In order to deliver genes for these two factors into 2-1, we developed a novel system that does not require drug selection and also employed a piggyBac-based system that permits introduction of multiple transgenes. We are now constructing 2-1-based new cell lines that are able to express both DAF and TM in the absence of α-Gal. The resulting cell line will be used for SCNT as donors to produce cloned piglets capable of expressing DAF and TM, but unable to express α-Gal.

Free Research Field

発生工学、分子生物学分野