Generation of a pig model for the diabetic nephropathy using gene editing technology

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K16014
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 42030:Animal life science-related
• Research Institution: Jichi Medical University (2020); The University of Tokushima (2019)
• Principal Investigator: Tanihara Fuminori 自治医科大学, 医学部, 准教授 (90754680)
• Project Period (FY): 2019-04-01 – 2021-03-31
• Keywords: ブタ / ゲノム編集 / CRISPR/Cas9 / エレクトロポレーション / 体外受精 / 糖尿病性腎症

Outline of Final Research Achievements

In this study, we aimed to generate diabetic nephropathy model pigs carrying both BDKRB2 and PDX1 genes modification from in vitro fertilized zygotes, which were prepared using sperm derived from PDX1 modified boar and introduced the CRISPR/Cas9 system targeting BDKRB2 gene by electroporation. Two gRNAs that efficiently modify the BDKRB2 gene were identified and used to generate gene-edited embryos, which were then transferred to embryonic pigs, but no offspring were obtained. In addition, we generated PDX1 monoallelic mutant pigs and observed their phenotypes related to diabetes. No health abnormalities were observed compared to wild-type pigs, and no difference was observed in the results of glucose tolerance tests. In pigs, monoallelic mutation of PDX1 gene alone is not sufficient to induce diabetes.

Free Research Field

動物発生工学

Academic Significance and Societal Importance of the Research Achievements

糖尿病患者は増加の一途を辿り、同時に問題となる糖尿病性腎症も含めてその病態解明および治療法の開発研究のため疾患モデルブタの担う役割は大きい。本研究では目的とする遺伝子改変ブタを得ることはできなかったが、本研究結果からPDX1遺伝子単独のヘテロノックアウトではブタでは糖尿病を誘発するには不十分であることが示唆された。PDX1遺伝子ヘテロノックアウトマウスでは膵臓β細胞からのインスリン分泌が低下することが知られているが、ブタでは表現型が異なることが示唆された。今後、糖尿病モデルブタの作出を目指す上で重要な知見が得られた。