Development of a novel mouse model of mosaic mutation-induced diseases caused by somatic mutations

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K06131
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43050:Genome biology-related
• Research Institution: Kazusa DNA Research Institute
• Principal Investigator: Nakayama Manabu 公益財団法人かずさDNA研究所, 先端研究開発部, 主任研究員 (30370927)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: ゲノムエンジニアリング / ゲノム改変技術 / 部位特異的組み換え酵素 / VCre/VloxP / SCre/SloxP / バイオテクノロジー

Outline of Final Research Achievements

Many mosaic mutations caused by somatic cell mutations have been observed in skin diseases. To produce a novel mouse model for these diseases and investigate their onset mechanisms, it is necessary to introduce gene knockouts or mutations into some but not all epidermal cells. Moreover, these cells should be fluorescently labelled for further analysis. In this study, by combining the Cre/loxP and CRISPR/Cas9 systems, we developed a new method through which strong Cre reactions reliably occur within single cells at low frequencies. We verified that this new method worked well by observation, under a fluorescence microscope, of reporter cells in which only one gene copy was knocked in (at specific locations of their genomes) and by next-generation sequencing analysis.

Free Research Field

機能ゲノム学

Academic Significance and Societal Importance of the Research Achievements

先天性角化症・魚鱗癬にはgain-of-functionである優性の体細胞突然変異が原因となり生じるモザイク疾患が多数存在し、それらのモザイク疾患を調べるためには、表皮細胞の一部だけ変異を導入かつ蛍光標識して解析する必要がある。また、皮膚細胞だけでなく、他の組織においても低頻度で細胞を標識するニーズは確実に存在している。本新規技術を他の組織の低頻度な細胞標識に用いることは比較的容易に達成できると考えられるので、多くの研究者が利用できるコアな技術となることが期待される。