Synthesis of a specific RNA-binding protein via the artificial evolution of Rf-PPR homologs

• Project/Area Number: 15K14626
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Science in genetics and breeding
• Research Institution: The University of Tokyo (2017); Nara Institute of Science and Technology (2015-2016)
• Principal Investigator: Fujii Sota 東京大学, 大学院農学生命科学研究科(農学部), 助教 (90716713)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 稔性回復遺伝子 / 人工進化 / RNA編集

Outline of Final Research Achievements

In the age of genome-editing, technologies that enable engineering of RNA expression is demanded. In this project, I aimed to establish such system by using the Rf-PPR protein, an RNA-binding protein with great diversity and ubiquitously present in angiosperm species. First of all, I established a system that visualizes the binding of a protein to an RNA sequence in E. coli. I have also performed functional characterization of an Rf-PPR homolog. The RFL2 was shown to be involved in the cleavage of the RNA transcript of the mitochondrially-encoded orf291.

Research Products

• [Journal Article] The Restorer-of-fertility-like 2 pentatricopeptide repeat protein and RNase P are required for the processing of mitochondrial orf291 RNA in Arabidopsis. (2016)
  • Author(s): Fujii S, Suzuki T, Giege; P, Higashiyama T, Koizuka N, Shikanai T.
  • Journal Title: Plant Journal Volume: 86 Issue: 6 Pages: 504-513
  • DOI: 10.1111/tpj.13185
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Calcium signalling mediates self-incompatibility response in the Brassicaceae (2015)
  • Author(s): Megumi Iwano, Kanae Ito, Sota Fujii, Mitsuru Kakita, Hiroko Asano-Shimosato, Motoko Igarashi, Pulla Kaothien-Nakayama, Tetsuyuki Entani, Asaka Kanatani, Masashi Takehisa, Masaki Tanaka, Kunihiko Komatsu, Hiroshi Shiba, Takeharu Nagai, Atsushi Miyawaki, Akira Isogai & Seiji Takayama
  • Journal Title: Nature Plants Volume: 1 Issue: 9 Pages: 15128-15128
  • DOI: 10.1038/nplants.2015.128
  • NAID: 120005716858
  • Peer Reviewed / Int'l Joint Research