Generation of African swine fever and highly pathogenic PRRS-resistant pig by genome editing of CD163

2019 Fiscal Year Final Research Report

• Project/Area Number: 17H03916
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: Kyushu University
• Principal Investigator: ONO ETSURO 九州大学, 医学研究院, 教授 (00160903)
• Co-Investigator(Kenkyū-buntansha): 大竹 正剛 静岡県畜産技術研究所, 中小家畜研究センター 養豚・養鶏, 上席研究員 (90605677)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: ゲノム編集 / 豚繁殖・呼吸障害症候群 / CD163 / アフリカ豚コレラ / マイクロミニピッグ

Outline of Final Research Achievements

Porcine reproductive and respiratory syndrome (PRRS) and African swine fever (ASF) are the most economically important disease of swine worldwide. We hypothesized that pigs with defective CD163 would be resistant to PRRS. Here, we have tried to generate PRRS-resistant pigs using CRISPR/Cas9 system by introducing amino acid mutations in the scavenger receptor cysteine-rich(SRCR5) domain of CD163 to keep the intact function and delete the virus receptor activity . Total 5 microminiature pig (MMP) embryonic fibroblast cell lines having indel in SRCR5 were established, which will be able to use somatic cell nuclear transfer for generating PRRS-resistant pigs. Cas9 protein, sgRNA and ssDNA for mutation were co-microinjected into the pronuclei of fertilized MMP embryos which were subsequently transplanted into pseudopregnant foster recipient pigs. It is, therefore, that the birth of PRRS-resistant MMP are expected.

Free Research Field

獣医微生物学

Academic Significance and Societal Importance of the Research Achievements

ゲノム編集による抗病性動物の作製方法の確立は、アフリカ豚コレラや豚繁殖・呼吸障害症候群をはじめとするワクチン開発が困難な感染症に対する新しい制圧法の開発に貢献する。現在、難治性感染症の病原体の多くが単球/マクロファージを初期感染細胞としているが、単球/マクロファージを準備することが困難で研究の進展を妨げている。胎仔線維芽細胞からのiPS細胞化、更に単球/マクロファージへの分化誘導法の確立は、感染症研究に新たな研究手法を提供する点において大きな意義がある。これらの研究は、将来の動物食料資源の確保と供給に貢献する。