title: A sensitive switch for visualizing natural gene silencing in single cells.
abstract: 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's 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.
authors.affiliation: School of Biological and Health Systems Engineering, Arizona State University, Tempe, AZ 85287, USA. karmella.haynes@asu.edu
authors.forename: Karmella A
authors.initials: KA
authors.lastname: Haynes
authors.forename: Francesca
authors.initials: F
authors.lastname: Ceroni
authors.forename: Daniel
authors.initials: D
authors.lastname: Flicker
authors.forename: Andrew
authors.initials: A
authors.lastname: Younger
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Silver
mesh_terms: D002460:Cell Line; D020868:Gene Silencing; D017930:Genes, Reporter; D005821:Genetic Techniques; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D035683:MicroRNAs / Q000235:genetics; D008957:Models, Genetic; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000235:genetics; D058615:Synthetic Biology
chemical_list: D008164:Luminescent Proteins; D035683:MicroRNAs; D011993:Recombinant Fusion Proteins