High-resolution Identification and Separation of Living Cell Types by Multiple microRNA-responsive Synthetic mRNAs

合成RNAを利用した生細胞の高精度な同定と分離. 京都大学プレスリリース. 2016-02-25.The precise identification and separation of living cell types is critical to both study cell function and prepare cells for medical applications. However, intracellular information to distinguish live cells remains largely inaccessible. Here, we develop a method for high-resolution identification and separation of cell types by quantifying multiple microRNA (miRNA) activities in live cell populations. We found that a set of miRNA-responsive, in vitro synthesized mRNAs identify a specific cell population as a sharp peak and clearly separate different cell types based on less than two-fold differences in miRNA activities. Increasing the number of miRNA-responsive mRNAs enhanced the capability for cell identification and separation, as we precisely and simultaneously distinguished different cell types with similar miRNA profiles. In addition, the set of synthetic mRNAs separated HeLa cells into subgroups, uncovering heterogeneity of the cells and the level of resolution achievable. Our method could identify target live cells and improve the efficiency of cell purification from heterogeneous populations

Numerical operations in living cells by programmable RNA devices

細胞内の複数のマイクロRNAを同時に検知して細胞を生きたまま精密に分けることに成功. 京都大学プレスリリース. 2019-08-30.Integrated bioengineering systems can make executable decisions according to the cell state. To sense the state, multiple biomarkers are detected and processed via logic gates with synthetic biological devices. However, numerical operations have not been achieved. Here, we show a design principle for messenger RNA (mRNA) devices that recapitulates intracellular information by multivariate calculations in single living cells. On the basis of this principle and the collected profiles of multiple microRNA activities, we demonstrate that rationally programmed mRNA sets classify living human cells and track their change during differentiation. Our mRNA devices automatically perform multivariate calculation and function as a decision-maker in response to dynamic intracellular changes in living cells

Effects of glucosamine on tooth pulpal nociceptive responses in the rat

AbstractBackground/purposed-Glucosamine hydrochloride (DGL) has a variety of biological activities and is noted as a nutritional supplement that is effective for improvement and care of various disorders, such as osteoarthritis and atherosclerosis. Although, it has been reported that DGL has a significant pain relief effect in treating osteoarthritis, little is known about its effect on dental pain. The applicability of DGL as a medicament to control pain in pulpalgia has not been reported. In this study, using an in vitro rat mandible-inferior alveolar nerve preparation (jaw-nerve preparation), the effect of DGL on nociceptive responses in the tooth pulpal nerve was examined.Materials and methodsThe effect of DGL on nociceptive responses for 20 male Wistar albino rats was evaluated using an in vitro jaw-nerve preparation. Bradykinin (BK), used as a chemical nociceptive stimulant, was applied near the exposed tooth pulp. Sixty seconds after BK application, the surface of the exposed pulp was treated with DGL solution or physiological saline (control).ResultsThe nerve firing rate was 2.06±0.21Hz (n=10) after 5 minutes of saline application, and 0.76±0.16Hz (n=10) after 5 minutes of DGL application. The DGL group showed significantly lower nerve firing rate than the control group.ConclusionBK-induced nociceptive responses were significantly suppressed by direct application of DGL. Our results suggest that DGL might have a pain relief effect in dental pain