The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo
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 extremel...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Public Library of Science
2014
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| Online Access: | http://hdl.handle.net/1721.1/89455 |
| _version_ | 1826212884893401088 |
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| author | Ruda, Vera M. Chandwani, Rohit Sehgal, Alfica Akinc, Akin Charisse, Klaus Tarakhovsky, Alexander Novobrantseva, Tatiana I. Kotelianski, Victor E. Bogorad, Roman |
| author2 | Koch Institute for Integrative Cancer Research at MIT |
| author_facet | Koch Institute for Integrative Cancer Research at MIT Ruda, Vera M. Chandwani, Rohit Sehgal, Alfica Akinc, Akin Charisse, Klaus Tarakhovsky, Alexander Novobrantseva, Tatiana I. Kotelianski, Victor E. Bogorad, Roman |
| author_sort | Ruda, Vera M. |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T15:39:41Z |
| format | Article |
| id | mit-1721.1/89455 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:39:41Z |
| publishDate | 2014 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/894552022-09-29T15:18:48Z The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo Ruda, Vera M. Chandwani, Rohit Sehgal, Alfica Akinc, Akin Charisse, Klaus Tarakhovsky, Alexander Novobrantseva, Tatiana I. Kotelianski, Victor E. Bogorad, Roman Koch Institute for Integrative Cancer Research at MIT Ruda, Vera M. Bogorad, Roman Kotelianski, Victor E. 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. Alnylam Pharmaceuticals (Firm) 2014-09-12T14:44:43Z 2014-09-12T14:44:43Z 2014-07 2013-12 Article http://purl.org/eprint/type/JournalArticle 1932-6203 http://hdl.handle.net/1721.1/89455 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. en_US http://dx.doi.org/10.1371/journal.pone.0101749 PLoS ONE Creative Commons Attribution http://creativecommons.org/licenses/by/4.0/ application/pdf Public Library of Science Public Library of Science |
| spellingShingle | Ruda, Vera M. Chandwani, Rohit Sehgal, Alfica Akinc, Akin Charisse, Klaus Tarakhovsky, Alexander Novobrantseva, Tatiana I. Kotelianski, Victor E. Bogorad, Roman The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo |
| title | The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo |
| title_full | The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo |
| title_fullStr | The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo |
| title_full_unstemmed | The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo |
| title_short | The Roles of Individual Mammalian Argonautes in RNA Interference In Vivo |
| title_sort | roles of individual mammalian argonautes in rna interference in vivo |
| url | http://hdl.handle.net/1721.1/89455 |
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