||MicroRNA-302 switch to identify and eliminate undifferentiated human pluripotent stem cells.
Parr, Callum J C ,
Katayama, Shota ,
Miki, Kenji ,
Kuang, Yi ,
Yoshida, Yoshinori ,
Morizane, Asuka ,
Takahashi, Jun ,
Yamanaka, ShinyaSaito, Hirohide
62016-09-09 , Springer Nature
iPS細胞を選択的に識別・分離・除去できるしくみを合成RNAを用いて構築. 京都大学プレスリリース. 2016-09-09.
The efficiency of pluripotent stem cell differentiation is highly variable, often resulting in heterogeneous populations that contain undifferentiated cells. Here we developed a sensitive, target-specific, and general method for removing undesired cells before transplantation. MicroRNA-302a-5p (miR-302a) is highly and specifically expressed in human pluripotent stem cells and gradually decreases to basal levels during differentiation. We synthesized a new RNA tool, miR-switch, as a live-cell reporter mRNA for miR-302a activity that can specifically detect human induced pluripotent stem cells (hiPSCs) down to a spiked level of 0.05% of hiPSCs in a heterogeneous population and can prevent teratoma formation in an in vivo tumorigenicity assay. Automated and selective hiPSC-elimination was achieved by controlling puromycin resistance using the miR-302a switch. Our system uniquely provides sensitive detection of pluripotent stem cells and partially differentiated cells. In addition to its ability to eliminate undifferentiated cells, miR-302a switch also holds great potential in investigating the dynamics of differentiation and/or reprograming of live-cells based on intracellular information.