Abstract

Efficient Detection and Purification of Cell Populations Using RNA Switches

Shin-Il Kim, Director, Division of Research & Development, aceRNA Technologies Co., Ltd.

Despite significant progress in improving the efficiency on a differentiation of specific cell types derived from pluripotent stem cells (PSCs), the isolation of target cells or the removal of unwanted cell population are required. Based on an endogenous microRNA (miRNA) activity in living cells, we developed an efficient method for purifying cells. We designed synthetic mRNAs encoding a reporter tagged with sequences targeted by cell-type specific miRNAs. These RNA switches control their translation levels by sensing miRNA activities. miR-208a-switch efficiently purified cardiomyocytes differentiated from human PSCs, and a switch encoding the apoptosis inducer Bim enriched for cardiomyocytes without cell sorting. This approach is generally applicable, as several miRNA-responsive switches precisely and efficiently isolated specific cell types differentiated from human PSCs. Thus, RNA switches can purify cell populations for which other isolation strategies are unavailable and could be useful to purify desired cell types for future therapeutic applications and regenerative medicine.