Journal article

Probing Transcription Factor Binding Activity and Downstream Gene Silencing in Living Cells with a DNA Nanoswitch

A Bertucci, J Guo, N Oppmann, A Glab, F Ricci, F Caruso, F Cavalieri

Nanoscale | The Royal Society of Chemistry | Published : 2018

Abstract

Transcription factor DNA binding activity is of pivotal importance in living systems because of its primary involvement in the regulation of genetic machinery. The analysis of transient expression levels of transcription factors in response to a certain cell status is a powerful means for investigating cellular dynamics at the biomolecular level. Herein, a DNA-based molecular switch that enables probing of transcription factor DNA binding activity is directly used in living cells. We demostrate that the DNA nanoswitch allows for dynamic fluorescence imaging of NF-?B and quantification of downstream gene silencing in real time. The present strategy is based on a functional DNA nanodevice that..

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Grants

Awarded by Australian Research council (ARC)


Awarded by Australian Government, Department of Education and Training


Awarded by ARC Centre of Excellence in Convergent Bio-Nano Science and Technology


Awarded by MSCA-RISE, NANOSUPREMI


Funding Acknowledgements

This work was supported by the Australian Research council (ARC) under the Future Fellowship (F.Cav., FT140100873) and Australian Laureate Fellowship (F.Car., FL120100030) schemes. A. B. thanks the Australian Government, Department of Education and Training for providing the Endeavour Research Fellowship (#5574) supporting postdoctoral research at The University of Melbourne. This research was also supported by the ARC Centre of Excellence in Convergent Bio-Nano Science and Technology (project number CE140100036) and MSCA-RISE, NANOSUPREMI PN: 690901. This work was performed in part at the Materials Characterisation and Fabrication Platform (MCFP) at The University of Melbourne and the Victorian Node of the Australian National Fabrication Facility (ANFF). We acknowledge Marcin Wojnilowicz and Paul Brannon for assistance with microscopy experiments. We also acknowledge Dr Jan Fric for helpful discussions.