A versatile and robust cell purification system with an RNA-only circuit composed of microRNA-responsive ON and OFF switches
| Autor: | Yoshihiko Fujita, Moe Hirosawa, Karin Hayashi, Takeshi Hatani, Yoshinori Yoshida, Takuya Yamamoto, Hirohide Saito |
|---|---|
| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Science Advances |
| ISSN: | 2375-2548 |
| Popis: | Human induced pluripotent stem cells (iPSCs) are promising cell resources for cell therapy and drug discovery. However, iPSC-derived differentiated cells are often heterogenous and need purification using a flow cytometer, which has high cost and time consumption for large-scale purification. MicroRNAs (miRNAs) can be used as cell selection markers, because their activity differs between cell types. Here, we show miRNA-responsive ON and OFF switch mRNAs for robust cell purification. The ON switch contains a miRNA-target sequence after the polyadenylate tail, triggering translational activation by sensing the target miRNA. By designing RNA-only circuits with miRNA-ON and -OFF switch mRNAs that encode a lethal ribonuclease, Barnase, and its inhibitor, Barstar, we efficiently purified specific cell types, including human iPSCs and differentiated cardiomyocytes, without flow cytometry. Synthetic mRNA circuits composed of ON and OFF switches provide a safe, versatile, and time-saving method to purify various cell types for biological and clinical applications. ２つの合成mRNAスイッチを活用した純度の高い細胞選別システムの開発. 京都大学プレスリリース. 2022-01-06. Synthetic gene circuits that improve stem cell quality. 京都大学プレスリリース. 2022-01-06. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|