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 Citations (Scopus)

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

Fingerprint

Gene Silencing
RNA Interference
MicroRNAs
Genes
Cells
Switches
Networks (circuits)
RNA
Gene expression
Chemical activation
Molecules
Sensors
Osteosarcoma
Gene Expression

Keywords

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

ASJC Scopus subject areas

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

Cite this

Haynes, K., Ceroni, F., Flicker, D., Younger, A., & Silver, P. A. (2012). A sensitive switch for visualizing natural gene silencing in single cells. ACS Synthetic Biology, 1(3), 99-106. https://doi.org/10.1021/sb3000035

A sensitive switch for visualizing natural gene silencing in single cells. / Haynes, Karmella; Ceroni, Francesca; Flicker, Daniel; Younger, Andrew; Silver, Pamela A.

In: ACS Synthetic Biology, Vol. 1, No. 3, 16.03.2012, p. 99-106.

Research output: Contribution to journalArticle

Haynes, K, Ceroni, F, Flicker, D, Younger, A & Silver, PA 2012, 'A sensitive switch for visualizing natural gene silencing in single cells', ACS Synthetic Biology, vol. 1, no. 3, pp. 99-106. https://doi.org/10.1021/sb3000035
Haynes, Karmella ; Ceroni, Francesca ; Flicker, Daniel ; Younger, Andrew ; Silver, Pamela A. / A sensitive switch for visualizing natural gene silencing in single cells. In: ACS Synthetic Biology. 2012 ; Vol. 1, No. 3. pp. 99-106.
@article{3e1174f3c63b442993f619f779cf4c50,
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.",
keywords = "Genetic switch, MiR-34, RNA interference, Synthetic repressor",
author = "Karmella Haynes and Francesca Ceroni and Daniel Flicker and Andrew Younger and Silver, {Pamela A.}",
year = "2012",
month = "3",
day = "16",
doi = "10.1021/sb3000035",
language = "English (US)",
volume = "1",
pages = "99--106",
journal = "ACS Synthetic Biology",
issn = "2161-5063",
publisher = "American Chemical Society",
number = "3",

}

TY - JOUR

T1 - A sensitive switch for visualizing natural gene silencing in single cells

AU - Haynes, Karmella

AU - Ceroni, Francesca

AU - Flicker, Daniel

AU - Younger, Andrew

AU - Silver, Pamela A.

PY - 2012/3/16

Y1 - 2012/3/16

N2 - 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.

AB - 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.

KW - Genetic switch

KW - MiR-34

KW - RNA interference

KW - Synthetic repressor

UR - http://www.scopus.com/inward/record.url?scp=84865304769&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84865304769&partnerID=8YFLogxK

U2 - 10.1021/sb3000035

DO - 10.1021/sb3000035

M3 - Article

C2 - 22530199

AN - SCOPUS:84865304769

VL - 1

SP - 99

EP - 106

JO - ACS Synthetic Biology

JF - ACS Synthetic Biology

SN - 2161-5063

IS - 3

ER -