Structural and functional analysis of RNA-guided effector nucleases

• Project/Area Number: 18H02384
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 43020:Structural biochemistry-related
• Research Institution: The University of Tokyo
• Principal Investigator: Nishimasu Hiroshi 東京大学, 先端科学技術研究センター, 教授 (00467044)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2020: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2018: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: RNA / タンパク質 / CRISPR / RNAサイレンシング / 立体構造 / piRNA / 結晶構造 / 結晶構造解析

Outline of Final Research Achievements

We determined the crystal structure of the PIWI protein from Drosophila melanogaster (Piwi). The structure revealed the molecular mechanism by which Piwi binds piRNA to protect germ cells from transposable elements. In addition, we determined the structures of Cas9 and Cas12 from various organisms, thereby improving our mechanistic understanding of diverse CRISPR-Cas enzymes and paving the way for further development of new genome-editing technologies.

Academic Significance and Societal Importance of the Research Achievements

原核生物のCRISPR-Cas獲得免疫機構および真核生物のRNAサイレンシング機構にかかわるRNA依存性ヌクレアーゼ複合体の構造機能解析を推進し、その多様な作動機構の一端を明らかにした。その結果、原核生物および真核生物のもつRNA依存性ヌクレアーゼの類似点および相違点が明らかになった。さらに、本研究の成果はゲノム編集やRNA干渉といった応用技術の開発基盤となることが期待される。

Research Products

• [Int'l Joint Research] Broad Institute(米国)
• [Int'l Joint Research] Broad Institute(米国)
• [Journal Article] Crystal structure of Drosophila Piwi. (2020)
  • Author(s): Yamaguchi, S., Oe, A., Nishida, KM., Yamashita, K., Kajiya, A., Hirano, S., Matsumoto, N., Dohmae, N., Ishitani, R., Saito, K., Siomi, H., Nishimasu, H., Siomi, MC., and Nureki, O.
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 858-858
  • DOI: 10.1038/s41467-020-14687-1
  • Peer Reviewed / Open Access
• [Journal Article] Structural basis for the promiscuous PAM recognition by Corynebacterium diphtheriae Cas9 (2019)
  • Author(s): Hirano Seiichi、Abudayyeh Omar O.、Gootenberg Jonathan S.、Horii Takuro、Ishitani Ryuichiro、Hatada Izuho、Zhang Feng、Nishimasu Hiroshi、Nureki Osamu
  • Journal Title: Nature Communications Volume: 10 Issue: 1 Pages: 1968-1968
  • DOI: 10.1038/s41467-019-09741-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genome editing in plants by engineered CRISPR-Cas9 recognizing NG PAM (2018)
  • Author(s): Endo Masaki、Mikami Masafumi、Endo Akira、Kaya Hidetaka、Itoh Takeshi、Nishimasu Hiroshi、Nureki Osamu、Toki Seiichi
  • Journal Title: Nature Plants Volume: 5 Issue: 1 Pages: 14-17
  • DOI: 10.1038/s41477-018-0321-8
  • Peer Reviewed / Open Access
• [Journal Article] Cap-specific terminal N6-methylation of RNA by an RNA polymerase II?associated methyltransferase (2018)
  • Author(s): Akichika Shinichiro、Hirano Seiichi、Shichino Yuichi、Suzuki Takeo、Nishimasu Hiroshi、Ishitani Ryuichiro、Sugita Ai、Hirose Yutaka、Iwasaki Shintaro、Nureki Osamu、Suzuki Tsutomu
  • Journal Title: Science Volume: 363 Issue: 363 Pages: 1-7
  • DOI: 10.1126/science.aav0080
  • Peer Reviewed / Open Access
• [Journal Article] Structural insights into cGAMP degradation by Ecto-nucleotide pyrophosphatase phosphodiesterase 1 (2018)
  • Author(s): Kato Kazuki、Nishimasu Hiroshi、Oikawa Daisuke、Hirano Seiichi、Hirano Hisato、Kasuya Go、Ishitani Ryuichiro、Tokunaga Fuminori、Nureki Osamu
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 4424-4424
  • DOI: 10.1038/s41467-018-06922-7
  • Peer Reviewed / Open Access
• [Journal Article] Engineered CRISPR-Cas9 nuclease with expanded targeting space (2018)
  • Author(s): Nishimasu Hiroshi、Shi Xi、Ishiguro Soh、Gao Linyi、Hirano Seiichi、Okazaki Sae、Noda Taichi、Abudayyeh Omar O.、Gootenberg Jonathan S.、Mori Hideto、Oura Seiya、Holmes Benjamin、Tanaka Mamoru、Seki Motoaki、Hirano Hisato、Aburatani Hiroyuki、Ishitani Ryuichiro、Ikawa Masahito、Yachie Nozomu、Zhang Feng、Nureki Osamu
  • Journal Title: Science Volume: 361 Issue: 6408 Pages: 1259-1262
  • DOI: 10.1126/science.aas9129
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 立体構造に基づく CRISPR-Cas9の分子改変 (2018)
  • Author(s): 西増 弘志
  • Organizer: MBSJ2018
  • Invited
• [Presentation] Engineered CRISPR-Cas9 recognizing a single guanine PAM (2018)
  • Author(s): Hiroshi Nishimasu
  • Organizer: 第91回日本生化学会大会
  • Invited
• [Remarks]
  • URL: https://www.s.u-tokyo.ac.jp/ja/press/2018/6021/