Generation of synthetic Plasmodium parasites.

• Project/Area Number: 19K22527
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 49:Pathology, infection/immunology, and related fields
• Research Institution: Osaka University (2020-2021); Tokyo Medical and Dental University (2019)
• Principal Investigator: IWANAGA SHIROH 大阪大学, 微生物病研究所, 教授 (20314510)
• Project Period (FY): 2019-06-28 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000); Fiscal Year 2020: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2019: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: マラリア原虫 / 人工細胞 / マラリア / 合成ゲノム / 人工染色体

Outline of Research at the Start

JC Venterらによる人工細菌の作製以後、いわゆる合成生物学が本格化し、人工真核生物の 合成が試みられている。しかし、ヒトやマウスの細胞の人工細胞合成は非常に困難であり 、これを打開するためには単純な真核生物で人工細胞を合成し、その成功に学ぶことが必須である。本研究は、ネズミマラリア原虫を用いて、実際に 細胞外で合成したゲノムスケール合成染色体を安定維持する人工マラリア原虫を作製する ことで、「マラリア原虫が人工細胞合成に適したモデル生物である」ことを実証すること を目的とする。

Outline of Final Research Achievements

Plasmodium parasites have extremely simple transcriptional regulation mechanism. In addition, the artificial chromosome technology can be used in the parasites. These biological and technical features may be useful for synthesizing the parasite artificially. In this study, to examine this idea, we attempted to synthesize the rodent malaria parasite, P. berghei. We developed a chromosome cleavage method by the CRISPR / Cas9 method, and prepared a recipient parasite for chromosome transplantation. Next, we created a shuttle artificial chromosome between Plasmodium parasite and Saccharomyces cerevisiae, and synthesized the small chromosome using it in S. cerevisiae. The synthesized small chromosome was successfully transplanted into the recipient parasites. These result suggested that the Plasmodium parasites is useful organism for synthesizing the eukaryotic cells.

Academic Significance and Societal Importance of the Research Achievements

本研究ではネズミマラリア原虫を用い、染色体切断法、出芽酵母内でのゲノム合成法、さらに合成ゲノムの原虫への移植法の開発に成功した。これまで真核生物においては人工細胞合成研究ではゲノム合成および移植の困難さから研究は停滞していた。よって、本研究の成果は真核生物における人工細胞合成に新たな進展を与えるものであり、その推進に貢献すると期待される。また、人工細胞は細胞機能の理解に新たな手法を提示するものであることから、本研究の成果はマラリア原虫の細胞機能の理解に貢献すると期待される。

Research Products

• [Journal Article] Insights and genetic features of extended-spectrum beta-lactamase producing Escherichia coli isolates from two hospitals in Ghana (2022)
  • Author(s): Mahazu Samiratu、Sato Wakana、Ayibieke Alafate、Prah Isaac、Hayashi Takaya、Suzuki Toshihiko、Iwanaga Shiroh、Ablordey Anthony、Saito Ryoichi
  • Journal Title: Scientific Reports Volume: 12 Issue: 1 Pages: 1843-1844
  • DOI: 10.1038/s41598-022-05869-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Chromosome splitting of Plasmodium berghei using the CRISPR/Cas9 system (2022)
  • Author(s): Addo-Gyan Daniel、Matsushita Haruka、Sora Enya、Nishi Tsubasa、Yuda Masao、Shinzawa Naoaki、Iwanaga Shiroh
  • Journal Title: PLOS ONE Volume: 17 Issue: 2 Pages: 0260176-0260176
  • DOI: 10.1371/journal.pone.0260176
  • Peer Reviewed / Open Access
• [Journal Article] Virulence Profiles of Diarrheagenic <i>Escherichia coli</i> Isolated from the Western Region of Ghana (2021)
  • Author(s): Prah Isaac、Ayibieke Alafate、Nguyen Thi Thu Huong、Iguchi Atsushi、Mahazu Samiratu、Sato Wakana、Hayashi Takaya、Yamaoka Shoji、Suzuki Toshihiko、Iwanaga Shiroh、Ablordey Anthony、Saito Ryoichi
  • Journal Title: Japanese Journal of Infectious Diseases Volume: 74 Issue: 2 Pages: 115-121
  • DOI: 10.7883/yoken.JJID.2020.356
  • NAID: 130008002375
  • ISSN: 1344-6304, 1884-2836
  • Year and Date: 2021-03-31
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Plasmodium falciparum RIFIN is a novel ligand for inhibitory immune receptor LILRB2. (2021)
  • Author(s): Sakoguchi A, Saito F, Hirayasu K, Shida K, Matsuoka S, Itagaki S, Nakai W, Kohyama M, Suenaga T, Iwanaga S, Horii T, Arase H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 548 Pages: 167-173
  • DOI: 10.1016/j.bbrc.2021.02.033
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Determining vector competence of Aedes aegypti from Ghana in transmitting dengue virus serotypes 1 and 2 (2021)
  • Author(s): Amoa-Bosompem Michael、Kobayashi Daisuke、Itokawa Kentaro、Murota Katsunori、Faizah Astri Nur、Azerigyik Faustus Akankperiwen、Hayashi Takaya、Ohashi Mitsuko、Bonney Joseph H. Kofi、Dadzie Samuel、Tran Cuong Chi、Tran Phong Vu、Fujita Ryosuke、Maekawa Yoshihide、Kasai Shinji、Yamaoka Shoji、Ohta Nobuo et al.
  • Journal Title: Parasites & Vectors Volume: 14 Issue: 1 Pages: 228-228
  • DOI: 10.1186/s13071-021-04728-z
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Emergence of oxacillinase-181 carbapenemase-producing diarrheagenic <i>Escherichia coli</i> in Ghana (2021)
  • Author(s): Prah Isaac、Ayibieke Alafate、Mahazu Samiratu、Sassa Chihiro Tani、Hayashi Takaya、Yamaoka Shoji、Suzuki Toshihiko、Iwanaga Shiroh、Ablordey Anthony、Saito Ryoichi
  • Journal Title: Emerging Microbes & Infections Volume: 10 Issue: 1 Pages: 865-873
  • DOI: 10.1080/22221751.2021.1920342
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Species composition and risk of transmission of some Aedes-borne arboviruses in some sites in Northern Ghana (2021)
  • Author(s): Joannides Joannitta、Dzodzomenyo Mawuli、Azerigyik Faustus、Agbosu Eudocia Esinam、Pratt Deborah、Nyarko Osei Joseph Harold、Pwalia Rebecca、Amlalo Godwin Kwame、Appawu Maxwell、Takashi Hayashi、Iwanaga Shiroh、Buchwald Andrea、Rochford Rosemary、Boakye Daniel、Koram Kwadwo、Bonney Kofi、Dadzie Samuel
  • Journal Title: PLOS ONE Volume: 16 Issue: 6 Pages: 0234675-0234675
  • DOI: 10.1371/journal.pone.0234675
  • Peer Reviewed / Open Access
• [Journal Article] Highly efficient CRISPR/Cas9 system in Plasmodium falciparum using Cas9-expressing parasites and a linear donor template (2021)
  • Author(s): Nishi Tsubasa、Shinzawa Naoaki、Yuda Masao、Iwanaga Shiroh
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 18501-18502
  • DOI: 10.1038/s41598-021-97984-z
  • Peer Reviewed / Open Access
• [Journal Article] Mechanisms of triggering malaria gametocytogenesis by AP2-G (2021)
  • Author(s): Yuda Masao、Kaneko Izumi、Murata Yuho、Iwanaga Shiroh、Nishi Tsubasa
  • Journal Title: Parasitology International Volume: 84 Pages: 102403-102403
  • DOI: 10.1016/j.parint.2021.102403
  • Peer Reviewed
• [Journal Article] Possible Dissemination of Escherichia coli Sequence Type 410 Closely Related to B4/H24RxC in Ghana (2021)
  • Author(s): Mahazu Samiratu、Prah Isaac、Ayibieke Alafate、Sato Wakana、Hayashi Takaya、Suzuki Toshihiko、Iwanaga Shiroh、Ablordey Anthony、Saito Ryoichi
  • Journal Title: Frontiers in Microbiology Volume: 12 Pages: 1-2
  • DOI: 10.3389/fmicb.2021.770130
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Screening for tick-borne and tick-associated viruses in ticks collected in Ghana (2021)
  • Author(s): Amoa-Bosompem Michael、Kobayashi Daisuke、Faizah Astri Nur、Kimura Shohei、Antwi Ama、Agbosu Esinam、Pratt Deborah、Ohashi Mitsuko、Bonney Joseph H. Kofi、Dadzie Samuel、Ejiri Hiroko、Ohta Nobuo、Sawabe Kyoko、Iwanaga Shiroh、Isawa Haruhiko
  • Journal Title: Archives of Virology Volume: 167 Issue: 1 Pages: 123-130
  • DOI: 10.1007/s00705-021-05296-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Rab5b-Associated Arf1 GTPase Regulates Export of N-Myristoylated Adenylate Kinase 2 From the Endoplasmic Reticulum in Plasmodium falciparum (2021)
  • Author(s): Taku Izumi、Hirai Tomohiro、Makiuchi Takashi、Shinzawa Naoaki、Iwanaga Shiroh、Annoura Takeshi、Nagamune Kisaburo、Nozaki Tomoyoshi、Saito-Nakano Yumiko
  • Journal Title: Frontiers in Cellular and Infection Microbiology Volume: 10 Pages: 610200-610200
  • DOI: 10.3389/fcimb.2020.610200
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Entomological Assessment of the Status and Risk of Mosquito-borne Arboviral Transmission in Ghana (2020)
  • Author(s): Amoa-Bosompem M, Kobayashi D, Murota K, Faizah A N、Itokawa K、Fujita R、Osei Joseph H N、Agbosu E、Pratt D、Kimura S、Kwofie Kofi D、Ohashi M、Bonney Joseph H. K、Dadzie S、Sasaki T、Ohta N、Isawa H、Sawabe K、Iwanaga S
  • Journal Title: Viruses Volume: 12 Issue: 2 Pages: 147-147
  • DOI: 10.3390/v12020147
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Improvement of CRISPR/Cas9 system by transfecting Cas9-expressing Plasmodium berghei with linear donor template (2020)
  • Author(s): Shinzawa Naoaki、Nishi Tsubasa、Hiyoshi Fumiya、Motooka Daisuke、Yuda Masao、Iwanaga Shiroh
  • Journal Title: Communications Biology Volume: 3 Issue: 1 Pages: 1-1
  • DOI: 10.1038/s42003-020-01138-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Establishment and characterization of a cell line from Ghanaian Aedes aegypti (Diptera: Culicidae) focusing on Aedes-borne flavivirus susceptibility (2020)
  • Author(s): Amoa-Bosompem Michael、Kobayashi Daisuke、Itokawa Kentaro、Faizah Astri Nur、Kuwata Ryusei、Dadzie Samuel、Hayashi Takaya、Yamaoka Shoji、Sawabe Kyoko、Iwanaga Shiroh、Isawa Haruhiko
  • Journal Title: In Vitro Cellular & Developmental Biology - Animal Volume: 56 Issue: 9 Pages: 792-798
  • DOI: 10.1007/s11626-020-00504-y
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Prevalence and Characterization of Carbapenem-Hydrolyzing Class D β-Lactamase-Producing Acinetobacter Isolates From Ghana (2020)
  • Author(s): Ayibieke Alafate、Kobayashi Ayumi、Suzuki Masato、Sato Wakana、Mahazu Samiratu、Prah Isaac、Mizoguchi Miyuki、Moriya Kyoji、Hayashi Takaya、Suzuki Toshihiko、Iwanaga Shiroh、Ablordey Anthony、Saito Ryoichi
  • Journal Title: Frontiers in Microbiology Volume: 11 Pages: 587398-587398
  • DOI: 10.3389/fmicb.2020.587398
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Female‐specific gene regulation in malaria parasites by an AP2‐family transcription factor (2019)
  • Author(s): Yuda Masao、Kaneko Izumi、Iwanaga Shiroh、Murata Yuho、Kato Tomomi
  • Journal Title: Molecular Microbiology Volume: 113 Issue: 1 Pages: 40-51
  • DOI: 10.1111/mmi.14334
  • Peer Reviewed / Open Access
• [Journal Article] CXCR4 regulates Plasmodium development in mouse and human hepatocytes (2019)
  • Author(s): Bando Hironori、Pradipta Ariel、Iwanaga Shiroh、Okamoto Toru、Okuzaki Daisuke、Tanaka Shun、Vega-Rodr?guez Joel、Lee Youngae、Ma Ji Su、Sakaguchi Naoya、Soga Akira、Fukumoto Shinya、Sasai Miwa、Matsuura Yoshiharu、Yuda Masao、Jacobs-Lorena Marcelo、Yamamoto Masahiro
  • Journal Title: Journal of Experimental Medicine Volume: 216 Issue: 8 Pages: 1733-1748
  • DOI: 10.1084/jem.20182227
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Antimalarial activity of vitamin D3 (VD3) does not result from VD3-induced antimicrobial agents including nitric oxide or cathelicidin (2019)
  • Author(s): Yamamoto Kiichi、Takahashi Kentaro、Ato Manabu、Iwanaga Shiroh、Ohta Nobuo
  • Journal Title: Experimental Parasitology Volume: 18 Pages: 30464-30468
  • DOI: 10.1016/j.exppara.2019.03.005
  • Peer Reviewed / Open Access
• [Presentation] 熱帯熱マラリア原虫Rab5bの結合タンパク質は N-ミリストイル化 adenylate kinase 2 を小胞体から選別輸送する (2021)
  • Author(s): 多久 和泉、平井 智浩、牧内 貴志、新澤 直明、岩永 史朗、案浦 健、 永宗 喜三郎、野崎 智義、中野 由美子
  • Organizer: 第90回日本寄生虫学会大会
• [Presentation] マラリア原虫生殖母体におけるRNA 結合型ジンクフィンガータンパク質による遺伝子発現制御機構の解析 (2021)
  • Author(s): 新澤 直明、関根 崇、平山 泰士、油田 正夫、岩永 史朗
  • Organizer: 第90回日本寄生虫学会大会
• [Presentation] 転写抑制因子AP2-FG2によるメス生殖母体の遺伝子制御 (2021)
  • Author(s): 西 翔、金子 伊澄、村田 優穂、岩永 史朗、油田 正夫
  • Organizer: 第90回日本寄生虫学会大会
• [Presentation] CRISPR/Cas9ゲノム編集法を用いた熱帯熱マラリア原虫の薬剤耐性遺伝子の再評価 (2021)
  • Author(s): 窪田 理恵、新澤 直明、岩永 史朗
  • Organizer: 第90回日本寄生虫学会大会
• [Presentation] Evaluation of the growth kinetics of Getah virus in a scope of mosquito cell lines (2021)
  • Author(s): Faustus Azerigyik, Daisuke Kobayashi, Michael Amoa-Bosompem, Astri Nur Faizah, Shiroh Iwanaga, Haruhiko Isawa
  • Organizer: 第73回日本衛生動物学会
• [Presentation] ガーナ産ネッタイシマカ由来新規培養細胞系の樹立と性状解析 (2021)
  • Author(s): 伊澤晴彦，マイケル・アモア-ボソムペム，小林大介，鍬田龍星，糸川健太郎， アシトリ・ヌー・ファイザー，岩永史朗
  • Organizer: 第73回日本衛生動物学会
• [Presentation] Screening for tick-borne and tick-associated viruses in ticks collected in Ghana (2021)
  • Author(s): Michael Amoa-Bosompem, Daisuke Kobayashi, Astri Nur Faizah, Hiroko Ejiri, Kyoko Sawabe, Shiroh Iwanaga, Haruhiko Isawa
  • Organizer: 第73回日本衛生動物学会
• [Presentation] Plasmodium falciparum RIFIN is a novel ligand for inhibitory immune receptor LILRB2 (2021)
  • Author(s): 迫口瑛史、岩永史朗、荒瀬尚
  • Organizer: 第85回日本インターフェロン・ サイトカイン学会学術集会
• [Presentation] フタトゲチマダニの胚発生における古典的トランスクリプトーム解析 (2020)
  • Author(s): Kofi Dazie KWOFIE, Emmanuel Pacia Helnandez, Samuel Dadzie, Danielle Ladzekpo、坪川大悟、三上房子、松林誠、岩永史朗、辻尚利、八田岳士
  • Organizer: 第72回日本衛生動物学会
• [Presentation] Virome analyses of Aedes aegypti mosquitoes collected in Ghana and their possible impact on arbovirus transmission (2020)
  • Author(s): Michael Amoa-Bosompem, Daisuke Kobayashi, Katsunori Murota, Astri Nur Faizah, Kentaro Itokawa, Mitsuko Ohashi, Toshinori Sasaki, Haruhiko Isawa, Kyoko Sawabe, Shiro Iwanaga
  • Organizer: 第72回日本衛生動物学会
• [Presentation] Transcriptional regulation of Plasmodium parasite (2020)
  • Author(s): Shiroh Iwanaga
  • Organizer: 第15回生命医科学ネットワークシンポジウム
  • Int'l Joint Research / Invited
• [Presentation] マラリア原虫有性ステージ分化におけるRNA結合型ジンクフィンガータンパク質ファミリーの解析 (2019)
  • Author(s): 平山 泰士、新澤 直明、油田 正夫、岩永 史朗
  • Organizer: 第79回 日本寄生虫学会東日本支部大会
• [Presentation] Female-specific gene regulation in malaria parasite by an AP2-family transcription factor. (2019)
  • Author(s): Shiroh Iwanaga
  • Organizer: The 18th Awaji International Forum on Infectiopn and Immunity
  • Invited