Development of cancer detection system using autonomous sequential reaction of nucleic acids

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K05819
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Analytical chemistry
• Research Institution: Kumamoto University
• Principal Investigator: Kitamura Yusuke 熊本大学, 大学院先端科学研究部(工), 助教 (80433019)
• Research Collaborator: Ihara Toshihiro
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 核酸プローブ / 細胞検出

Outline of Final Research Achievements

We tried to construct the detection system of target cells with high sensitivity based on the signal amplification triggered by a protein highly expressed on the membrane of target cells. A nucleic acid aptamer with arbitrary tag sequence was used as an initiator of the amplification reaction. A single stranded DNA which can work as a trigger for DNA circuit of signal amplification reaction was successfully released from a duplex by the strand exchange with the tag tethered with aptamer bound on the target cell surface. Specific initiation of DNA circuit by the trigger was confirmed in vitro. Then, we tried to detect the target cell by the initiation of DNA circuit using the trigger specifically released by the tag on the target cell. However, signal amplification was not efficient. The optimization of condition of DNA circuit should be needed for obtaining enough signal to detect target cells.

Free Research Field

分析化学、核酸化学

Academic Significance and Societal Importance of the Research Achievements

現在までのところ、DNAサーキットによるシグナル増幅は不十分ではあるが、条件を最適化できた際には、標的細胞を現行法のように顕微鏡で直接検出する必要なく、標的細胞を含む溶液の発光にて検出することが可能となる。標的細胞に任意のタグを提示し、鎖交換反応を自在に起こすことができたため、これに伴って放出された任意の配列の核酸を開始剤とする様々な核酸の自発的連鎖反応を誘起可能であることがわかった。