Trial testing for generating gene-modified Schistosoma japonicum by the use of retroviruses

• Project/Area Number: 26670198
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Parasitology (including sanitary zoology)
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Ohta Nobuo 東京医科歯科大学, 医歯(薬)学総合研究科, 教授 (10143611)
• Co-Investigator(Kenkyū-buntansha): KUMAGAI Takashi 東京医科歯科大学, 大学院医歯学総合研究科, 助教 (40369054)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 日本住血吸虫 / 遺伝子改変 / レンチウイルス / GFPタンパク質 / 遺伝子導入 / Anchor-PCR法 / 蛍光発現住血吸虫

Outline of Final Research Achievements

Since schistosome parasites use intermediate snail hosts for their life cycle, gene manipulation techniques is not easy to use for schistosome parasites. To overcome the difficulty, an approach using Letivirus was considered. In our trial, it was tested to insert GFP gene into the parasite genome together with Lentivirus. After then, it was possible to detect mRNA of GFP gene, and it was also confirmed that GFP gene was inserted into schistosome genome. There was no flame-shift of the inserted gene. However, we could not detect expression of GFP in the parasite, suggesting the presence of inhibition mechanism for protein expression from mRNA. ALthough protein expression was not available in our new system, it might have been possible to construct gene-knockdown system by inducing mRNA with inhibitory functions.

Research Products

• [Journal Article] Immunogenicity and anti-fecundity effect of nanoparticle coated glutathione S-transferase (SjGST) DNA vaccine against murine Schistosoma japonicum infection. (2015)
  • Author(s): Mbanefo EC, Kumagai T, Kodama Y, Kurosaki T, Furushima-Shimogawara R, Cherif MS, Mizukami S, Kikuchi M, Huy NT, Ohta N, Sasaki H, Hirayama K.
  • Journal Title: Parasitol Int. Volume: 64 Issue: 4 Pages: 24-31
  • DOI: 10.1016/j.parint.2015.01.005
  • Peer Reviewed / Open Access
• [Presentation] Female-biased miRNA production through the extracellular vesicles induced by the erythrocyte uptake in the adult worms of Schistosoma japonicum. (2016)
  • Author(s): Kumagai T, Ichimura K, Yamabe M, Shimogawara R, Ohta N.
  • Organizer: Annual meeting of US-Japan Cooperative Medical Science Program
  • Place of Presentation: Washington DC, USA
  • Year and Date: 2016-01-13
  • Int'l Joint Research
• [Presentation] 赤血球の摂取、及びペアリングによって分泌される日本住血吸虫の細胞外小胞内mRNA. (2015)
  • Author(s): 熊谷貴、市村浩一郎、山邊将史、下河原理江子、太田伸生
  • Organizer: 第38回日本分子生物学会 第88回日本生化学会合同会議
  • Place of Presentation: 兵庫県神戸市、神戸国際会議場
  • Year and Date: 2015-12-01
• [Book] 寄生虫学研究 材料と方法 2014 (2014)
  • Author(s): 熊谷 貴、下河原 理江子、山邊 将史、太田伸生
  • Total Pages: 7
  • Publisher: 三恵社