A sensitive switch for visualizing natural gene silencing in single cells

Karmella Haynes, Francesca Ceroni, Daniel Flicker, Andrew Younger, Pamela A. Silver

Research output: Contribution to journalArticle

7 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)99-106
Number of pages8
JournalACS Synthetic Biology
Volume1
Issue number3
DOIs
StatePublished - Mar 16 2012

Keywords

  • Genetic switch
  • MiR-34
  • RNA interference
  • Synthetic repressor

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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