Development of genetic code restoration by using artificial RNA editing

2015 Fiscal Year Final Research Report

• Project/Area Number: 26670167
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Human genetics
• Research Institution: Japan Advanced Institute of Science and Technology
• Principal Investigator: TSUKAHARA Toshifumi 北陸先端科学技術大学院大学, マテリアルサイエンス研究科, 教授 (60207339)
• Co-Investigator(Renkei-kenkyūsha): TATENO MASARU 兵庫県立大学, 院・生命理学研究科, 教授 (40291926) ; SUZUKI HITOSHI 北陸先端科学技術大学院大学, マテリアルサイエンス研究科, 特任助教 (00447690)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: Deaminase / RNA editing / MS2 / 活性ドメイン / guide RNA / 遺伝子修復

Outline of Final Research Achievements

To imitate RNA editing system for artificial and site-specific deamination of nucleic acid toward base-substitutions in vivo, we connected the active domain of ADAR1 with a guide RNA by using MS2 system. The nonsense mutated EGFP was used as a reporter gene. These 3 genes were transiently transfected into HEK293 cells at once. We observed green fluorescent cells in which parts of mutated EGFP mRNAs were restored in vivo. The genetic code restoration was confirmed by RT-PCR-RFLP and sequencing, respectively. We also compared efficiencies of RNA restorations by the site-specific deamination among ADAR family isoforms.

Free Research Field

生化学・分子生物学