The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo
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Ruda-2014-The roles of individ.pdf
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Author(s)
Ruda, Vera M.
Chandwani, Rohit
Sehgal, Alfica
Akinc, Akin
Charisse, Klaus
Tarakhovsky, Alexander
Novobrantseva, Tatiana I.
Kotelianski, Victor E.
Bogorad, Roman
Date Issued
July 2014
Journal
PLoS ONE
Publisher
Public Library of Science
Citation
Ruda, Vera M., Rohit Chandwani, Alfica Sehgal, Roman L. Bogorad, Akin Akinc, Klaus Charisse, Alexander Tarakhovsky, Tatiana I. Novobrantseva, and Victor Koteliansky. “The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo.” Edited by Alexandre H. Kihara. PLoS ONE 9, no. 7 (July 3, 2014): e101749.
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Final published version
Abstract
Argonaute 2 (Ago2) is the only mammalian Ago protein capable of mRNA cleavage. It has been reported that the activity of the short interfering RNA targeting coding sequence (CDS), but not 3′ untranslated region (3′UTR) of an mRNA, is solely dependent on Ago2 in vitro. These studies utilized extremely high doses of siRNAs and overexpressed Ago proteins, as well as were directed at various highly expressed reporter transgenes. Here we report the effect of Ago2 in vivo on targeted knockdown of several endogenous genes by siRNAs, targeting both CDS and 3′UTR. We show that siRNAs targeting CDS lose their activity in the absence of Ago2, whereas both Ago1 and Ago3 proteins contribute to residual 3′UTR-targeted siRNA-mediated knockdown observed in the absence of Ago2 in mouse liver. Our results provide mechanistic insight into two components mediating RNAi under physiological conditions: mRNA cleavage dependent and independent. In addition our results contribute a novel consideration for designing most efficacious siRNA molecules with the preference given to 3′UTR targeting as to harness the activity of several Ago proteins.
MIT Department
Koch Institute for Integrative Cancer Research at MIT
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DOI of Published Version
https://doi.org/10.1371/journal.pone.0101749
