The development for the stable high expression of numerous transgenes.

2018 Fiscal Year Final Research Report

• Project/Area Number: 16H05401
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General surgery
• Research Institution: Osaka University
• Principal Investigator: Miyagawa Shuji 大阪大学, 医学系研究科, 准教授 (90273648)
• Co-Investigator(Kenkyū-buntansha): 長嶋 比呂志 明治大学, 農学部, 専任教授 (50318664)
• Research Collaborator: Ikawa Masahito
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: special construct / pig ROSA26 locus / CD200 / CL-SP-D / CD31 / PQA-18

Outline of Final Research Achievements

We have produced special construct genes. The expression of NCTDM was not efficient, then separated to C1DM & TM. HLA-Ev(147)-2A-beta2m was also produced. In addition, we produced PI-anchored CD47, PI-anchored TFPI. A project of pig ROSA26 locus was started. We demonstrated the targeted knock-in of a gene cassette in this locus. Three guide RNA-sequences were designed, using CRISPR/Cas9 system, and the cleavage efficiency of each plasmid was assessed.
We studied novel strategies for suppressing macrophages and neutrophil-mediated xenogeneic rejection using pig ROSA26 locus. Moreover, we focused on the effect of new immunosuppressants on monocyte/macrophages, such as a PQA-18. PQA-18 showed the potential function on macrophage-mediated cytotoxicity, as an immunosuppressant for xenotransplantation.

Free Research Field

異種移植

Academic Significance and Societal Importance of the Research Achievements

今回の我々の研究は、国際的にも独自性が高いものであり、遺伝子改変ブタ作りに有効である。また、ROSA26の座に遺伝子をknockinさせる方法を確立させる事も有効である。更に、新たに異種移植に有効な分子を数々検定し、また免疫抑制剤を研究する事も、この分野に非常に有効と考えられる。
我々がこれらの遺伝子改変ブタを作出できなければ、外国の遺伝子改変ブタを使う事になる。これに伴い、関連する分野も含めて日本のこの分野での立ち遅れが必至となる。逆に、成功すれば、アジアを中心に、アメリカ、欧州への輸出が可能になり、これまで人の死を前提としていた移植医療を根本的に変える事になる。