Development of intracellular binary counter for recording the gene expression change

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K14603
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Tumor biology
• Research Institution: Hokkaido University
• Principal Investigator: Onodera Yasuhito 北海道大学, 医学研究院, 講師 (90435561)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 遺伝子発現制御

Outline of Final Research Achievements

In this study, we sought to develop a set of genetic constructions which records the change of the expression (ON/OFF) of the gene of interest in each cell. In order to record as many times of the expression change as possible, we aimed for the construction of the binary counter. For the proper function, these constructs require the single copy integration into the target cell genome, which is currently achieved by genome editing followed by single cell cloning. In these processes, however, heterogeneity of the cells, which is also a primary subject of this study, will be inevitably compromised. In particular, for the analyses of cancer cells, loss of heterogeneity may lead to contradictory results. Therefore, we also sought to establish a novel genetic markers by which single copy integration of multiple genetic constructs can be easily achieved.

Free Research Field

分子生物学、分子腫瘍学

Academic Significance and Societal Importance of the Research Achievements

現状では、遺伝子発現の増減を解析する手法は確立されているものの、その可塑性についての解析法は非常に限定的であり、単一細胞レベルではさらに困難である。がん細胞のような「不均一性」を特徴とする細胞集団の場合、解析対象とする細胞の遺伝子発現変化を長期間追跡することはほぼ不可能で有る。これを定量的に解析する手法が確立されれば、可塑性の調節を原理としたがん治療などが可能になると考えられる。また、本研究で必須となる遺伝子導入後の選択に関する技術は、今回行ったようなゲノム編集を要する遺伝子導入のみならず、一般に広く用いられている遺伝子導入後の安定発現株の樹立においても非常に有用なものである。