A Sensitive Switch for Visualizing Natural Gene Silencing in Single Cells
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- 作者：Karmella A. Haynes ; Francesca Ceroni ; Daniel Flicker ; Andrew Younger ; Pamela A. Silver
- 刊名：ACS Synthetic Biology
- 出版年：2012
- 出版时间：March 16, 2012
- 年：2012
- 卷：1
- 期：3
- 页码：99-106
- 全文大小：434K
- 年卷期：v.1,no.3(March 16, 2012)
- ISSN：2161-5063
文摘
RNA interference is a natural gene expression silencing system that appears throughout the tree of life. As the list of cellular processes linked to RNAi grows, so does the demand for tools to accurately measure RNAi dynamics in living cells. We engineered a synthetic RNAi sensor that converts this negative regulatory signal into a positive output in living mammalian cells, thereby allowing increased sensitivity and activation. Furthermore, the circuit鈥檚 modular design allows potentially any microRNA of interest to be detected. We demonstrated that the circuit responds to an artificial microRNA and becomes activated when the RNAi target is replaced by a natural microRNA target (miR-34) in U2OS osteosarcoma cells. Our studies extend the application of rationally designed synthetic switches to RNAi, providing a sensitive way to visualize the dynamics of RNAi activity rather than just the presence of miRNA molecules.