RNAi revised - Target mRNA-dependent enhancement of gene silencing

Simon Dornseifer; Sarah Willkomm; Rosel Kretschmer Kazemi Far; Janine Liebschwager; Foteini Beltsiou; Kirsten Frank; Sandra D. Laufer; Thomas Martinetz; Georg Sczakiel; Jens Christian Claussen; Tobias Restle

doi:10.1093/nar/gkv1200

RNAi revised - Target mRNA-dependent enhancement of gene silencing

Simon Dornseifer, Sarah Willkomm, Rosel Kretschmer Kazemi Far, Janine Liebschwager, Foteini Beltsiou, Kirsten Frank, Sandra D. Laufer, Thomas Martinetz, Georg Sczakiel, Jens Christian Claussen, Tobias Restle^*

^*Corresponding author for this work

College of Engineering and Physical Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

The discovery of RNA interference (RNAi) gave rise to the development of new nucleic acid-based technologies as powerful investigational tools and potential therapeutics. Mechanistic key details of RNAi in humans need to be deciphered yet, before such approaches take root in biomedicine and molecular therapy. We developed and validated an in silico-based model of siRNA-mediated RNAi in human cells in order to link in vitro-derived pre-steady state kinetic data with a quantitative and time-resolved understanding of RNAi on the cellular level. The observation that product release by Argonaute 2 is accelerated in the presence of an excess of target RNA in vitro inspired us to suggest an associative mechanism for the RNA slicer reaction where incoming target mRNAs actively promote dissociation of cleaved mRNA fragments. This novel associative model is compatible with highmultiple turnover rates of RNAibased gene silencing in living cells and accounts for target mRNA concentration-dependent enhancement of the RNAi machinery.

Original language	English
Pages (from-to)	10623-10632
Number of pages	10
Journal	Nucleic Acids Research
Volume	43
Issue number	22
Early online date	17 Nov 2015
DOIs	https://doi.org/10.1093/nar/gkv1200
Publication status	Published - 15 Dec 2015

Bibliographical note

© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact [email protected]

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10.1093/nar/gkv1200Licence: CC BY-NC 3.0

RNAi revised - target mRNA-dependent enhancement of gene silencing
© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact [email protected]
Final published version, 10.6 MBLicence: CC BY-NC 3.0

Cite this

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abstract = "The discovery of RNA interference (RNAi) gave rise to the development of new nucleic acid-based technologies as powerful investigational tools and potential therapeutics. Mechanistic key details of RNAi in humans need to be deciphered yet, before such approaches take root in biomedicine and molecular therapy. We developed and validated an in silico-based model of siRNA-mediated RNAi in human cells in order to link in vitro-derived pre-steady state kinetic data with a quantitative and time-resolved understanding of RNAi on the cellular level. The observation that product release by Argonaute 2 is accelerated in the presence of an excess of target RNA in vitro inspired us to suggest an associative mechanism for the RNA slicer reaction where incoming target mRNAs actively promote dissociation of cleaved mRNA fragments. This novel associative model is compatible with highmultiple turnover rates of RNAibased gene silencing in living cells and accounts for target mRNA concentration-dependent enhancement of the RNAi machinery.",

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RNAi revised - Target mRNA-dependent enhancement of gene silencing

Abstract

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