Development of mutator Plasmodium falciparum and its application for mutants generation.

• Project/Area Number: 15K19089
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Parasitology (including sanitary zoology)
• Research Institution: Tokyo Women's Medical University
• Principal Investigator: Honma Hajime 東京女子医科大学, 医学部, 助教 (10617468)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: マラリア / Plasmodium falciparum / MSH2 / ミスマッチDNA修復 / CRISPR/Cas9 / ミューテーター / ミスマッチ / CRISPR/Cas / MLH1 / Plasmodium

Outline of Final Research Achievements

MSH2-1 is thought to be one of the major DNA mismatch repair proteins of Plasmodium species. The MSH2-1 mutant of P. falciparum 3D7 was generated by substituting the 513rd proline to threonine with CRISPR/Cas9 based gene editing. The parent strain 3D7 and the MSH2-1 mutant were maintained by the continuous in vitro culture for around 200 days, and then spontaneous mutations were genome-widely evaluated by high-throughput sequencing. Increases of insertion/deletion mutations in microsatellites and structural variants in subtelomeric regions were observed in the MSH2-1 mutant.

Research Products

• [Journal Article] Draft Genome Sequence of Plasmodium gonderi, a Malaria Parasite of African Old World Monkeys (2017)
  • Author(s): Honma H, Kawai S, Motooka D, Nakamura S, Tougan T, Horii T, Arisue N.
  • Journal Title: Genome Announc. Volume: 5 Issue: 28
  • DOI: 10.1128/genomea.00612-17
  • Peer Reviewed / Open Access
• [Journal Article] Mutation tendency of mutator Plasmodium berghei with proofreading-deficient DNA polymerase δ. (2016)
  • Author(s): Honma H, Niikura M, Kobayashi F, Horii T, Mita T, Endo H, Hirai M
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 36971-36971
  • DOI: 10.1038/srep36971
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] ディープシーケンスによるマラリア原虫のアトバコン耐性に関わる点変異の解析 (2018)
  • Author(s): 彦坂健児、馮雪、本間一、松崎素道、元岡大祐、中村昇太、野呂瀬一美、北潔
  • Organizer: 第87回日本寄生虫学会大会
• [Presentation] 熱帯熱マラリア原虫のDNA修復タンパク質MSH2-1変異体の作製と性状解析 (2018)
  • Author(s): 本間一、高橋延之、杉下智彦
  • Organizer: 第87回日本寄生虫学会
• [Presentation] Whole-genome sequencing of a Plasmodium gonderi and evolutionary relationship of malaria parasites (2017)
  • Author(s): Nobuko Arisue, Hajime Honma, Satoru Kawai, Takahiro Tougan, Toshihiro Horii
  • Organizer: International Union of Microbiological Societies IUMS2017: 15th International Congress of Bacteriology & Applied Microbiology
  • Int'l Joint Research
• [Presentation] マラリア原虫の遺伝子点変異によるアトバコン耐性獲得様式の解明 (2016)
  • Author(s): 彦坂健児, 本間一, 松崎素道, 野呂瀬一美, 北潔
  • Organizer: 第85回日本寄生虫学会大会
  • Place of Presentation: 宮崎市民プラザ
  • Year and Date: 2016-03-19
• [Presentation] サルマラリア原虫Plasmodium gonderiのゲノム解析－マラリア原虫の系統関係解明に向けて－ (2015)
  • Author(s): 有末伸子, 本間一, 川合覚, 東岸任弘, 田邊和裄, 堀井俊宏
  • Organizer: 第23回分子寄生虫学ワークショップ／ 第13回分子寄生虫・マラリア研究フォーラム合同大会
  • Place of Presentation: 帯広畜産大学・原虫病研究センター PKホール
  • Year and Date: 2015-08-30
• [Remarks] マラリア原虫のゲノム解析～マラリア感染制御への可能性～
  • URL: http://www.twmu.ac.jp/univ/news/?kbn=1&ym=201612
• [Remarks] 薬剤耐性マラリア対策の画期的研究ツールを開発～ミューテーターでマラリアの変異を追う～
  • URL: http://www.juntendo.ac.jp/news/20161116-01.html