Dynamics of mtDNA in neuron

2018 Fiscal Year Final Research Report

• Project/Area Number: 17H07369
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: General anatomy (including histology/embryology)
• Research Institution: Institute of Physical and Chemical Research
• Principal Investigator: Komatsubara Akira 国立研究開発法人理化学研究所, 生命機能科学研究センター, 研究員 (70802792)
• Project Period (FY): 2017-08-25 – 2019-03-31
• Keywords: dCas9 / MS2

Outline of Final Research Achievements

To develop a technique for labeling mitochondrial genomes in living cells, we examined the extent to which telomeric regions stain using the CRISPR / Cas9 system, which is frequently used to label specific regions of the nuclear genome. When 16 cells of MS2 sequence were connected to sgRNA targeting telomeric region, MCP-GFP in which GFP was connected to MCP which recognizes MS2 sequence, and dCas9 were expressed in one cell, GFP was in the nucleus. It was confirmed that it was aggregated. The cells were fixed for verification and immunostaining of telomeric regions showed co-localization with MCP-GFP. From the above, it is thought that specific regions of nucleic acid can be stained in living cells by the method using dCas9 and MS2.

Free Research Field

クロマチン構造

Academic Significance and Societal Importance of the Research Achievements

ミトコンドリアは核ゲノムより転写翻訳されたタンパク質だけでなく、ミトコンドリアゲノム（mtDNA)から発現するタンパク質にも依存して機能を維持しているため、本研究ではmtDNAの挙動を調べるためにCRISRP/Cas9の技術を用いてゲノム特定部位を標識する実験を行った。生細胞内におけるゲノム特定領域の可視化技術は今後ミトコンドリアの機能維持の機構を知る上で重要な意味を持つと考えられる。