Identification and separation of living cells based on synthetic RNA circuits detecting intracellular information

2015 Fiscal Year Final Research Report

• Project/Area Number: 25870355
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General medical chemistry; System genome science
• Research Institution: The University of Tokyo (2014-2015); Kyoto University (2013)
• Principal Investigator: ENDO Kei 東京大学, 新領域創成科学研究科, 助教 (40626074)
• Co-Investigator(Renkei-kenkyūsha): SAITO Hirohide 京都大学, iPS細胞研究所, 教授 (20423014)
• Research Collaborator: HAYASHI Karin
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 細胞分画法 / mRNA / マイクロRNA / 人口遺伝子回路 / 合成生物学 / 再生医療

Outline of Final Research Achievements

Methods for identifying the cell types are critical not only to analysis of specific tissues and cells, but also to preparation of particular cells for therapeutic purposes. Marker proteins on the surface of living cells have been investigated to identify target cells. However, it is still hard to find a specific set of antibodies that determines a variety of cell types. In addition, possible combinations of binary classification are quite limited.
In this project, we developed a method that enables to distinguish living cells based on intracellular markers in a quantitative manner. We focused on the activity of microRNAs, which are short, non-coding RNAs expressed in a cell. We designed and synthesized microRNA-responsive mRNAs that produce fluorescent proteins as a reporter, and transfected them directly to target cells. With the synthetic mRNAs, we succeeded in isolating multiple cell types based on subtle (less than 2-fold) difference in the microRNA activity.

Free Research Field

分子生物学, 合成生物学, RNA医工学