Attempts to perform genome modification towards donor cells for improving SCNT efficiency in pigs

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K06372
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 42010:Animal production science-related
• Research Institution: National Center for Child Health and Development (2021); Kagoshima University (2019-2020)
• Principal Investigator: Sato Masahiro 国立研究開発法人国立成育医療研究センター, ゲノム医療研究部, 共同研究員 (30287099)
• Co-Investigator(Kenkyū-buntansha): 三好 和睦 鹿児島大学, 農水産獣医学域農学系, 教授 (70363611)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 体細胞核移植 / クローンブタ / 遺伝子改変 / DNAメチル化 / ゲノム編集 / DNAメチル化転移酵素1遺伝子 / Oct-3/4 / EGFP

Outline of Final Research Achievements

For enhancing the production efficiency of cloned piglets derived from somatic cell nuclear transfer, disruption of DNA methyltransferase 1 (DNMT1) gene, which is thought to be important for DNA methylation, in the donor porcine fibroblasts was attempted using CRISPR/Cas9 system. Notably, in the genome of this donor cell an expression unit carrying Oct-3/4 promoter linked to enhanced green fluorescent protein (EGFP) cDNA has already been inserted. SCNT using the donor cells with mutated DNMT1 gene resulted in an increased developmental rate to blastocysts about 2-folds as well as increased expression of EGFP. These results indicate an importance of inhibiting DNA methylation to improve SCNT.

Free Research Field

発生工学

Academic Significance and Societal Importance of the Research Achievements

本研究は、医学や農学分野で重要なブタの遺伝子改変を通じ、ヒトの健康・福祉に貢献する遺伝子改変クローンブタを効率的に作製することを目指す。そのカギとなるのが、ヒストンのDNAメチル化問題であり、今回、その是非をゲノム編集法と体細胞核移植（SCNT）実験により解明した（学術的意義）。これにより、クローンブタ作製は加速。その効果は医学や農林水産業の振興・発展に貢献すると考えられる（社会的意義）。