Generation of PRRS-resistant pigs using CRISPR/Cas9 system

2016 Fiscal Year Final Research Report

• Project/Area Number: 26292151
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: Kyushu University
• Principal Investigator: Ono Etsuro 九州大学, 医学(系)研究科(研究院), 教授 (00160903)
• Co-Investigator(Kenkyū-buntansha): 河原崎 達雄 東海大学, 農学部, 教授 (70500247) ; 柴田 昌利 静岡県畜産技術研究所, その他部局等, その他 (70501443)
• Co-Investigator(Renkei-kenkyūsha): IWAMORI NAOKI 九州大学, 大学院農学研究院, 准教授 (70647362) ; TOMIOKA YUKIKO 鳥取大学, 農学部, 准教授 (50374674) ; FUJIMOTO YOSHIKAZU 九州大学, 大学院医学研究院, 助教 (20613631)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 豚繁殖・呼吸障害症候群 / ゲノム編集 / 抗病性動物 / PRRS

Outline of Final Research Achievements

Porcine reproductive and respiratory syndrome (PRRS) is the most economically important disease of swine worldwide. Vaccines have been unable to control the disease. It has been suggested that CD163 and/or Siglec 1 are the receptors for entry of porcine reproductive and respiratory syndrome virus (PRRSV) into cells. Thus, we hypothesized that pigs with defective CD163 and/or Siglec 1 would be resistant to PRRSV. Here, we have tried to generate PRRS-resistant pigs using CRISPR/Cas9 system by introducing mutations in the CD163 and Siglec 1 genes to secrete a soluble form of each molecule. Total 11 piglets were born from foster pigs transferred zygotes injected with each sgRNA and Cas9 and an adenine insertion was detected in the Siglec 1 gene in one of them. Unfortunately, the piglet exhibited a mosaic genotype. In addition, somatic cell nuclear transfer was performed using porcine fibroblasts possessing the CD163 gene edited by CRISPR/Cas9 system.

Free Research Field

獣医微生物学