title: The roles of individual mammalian argonautes in RNA interference in vivo.
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.
authors.affiliation: Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Vera M
authors.initials: VM
authors.lastname: Ruda
authors.affiliation: Laboratory of Immune Cell Epigenetics and Signaling, Rockefeller University, New York, New York, United States of America.
authors.forename: Rohit
authors.initials: R
authors.lastname: Chandwani
authors.affiliation: Alnylam Pharmaceuticals, Cambridge, Massachusetts, United States of America.
authors.forename: Alfica
authors.initials: A
authors.lastname: Sehgal
authors.affiliation: Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Roman L
authors.initials: RL
authors.lastname: Bogorad
authors.affiliation: Alnylam Pharmaceuticals, Cambridge, Massachusetts, United States of America.
authors.forename: Akin
authors.initials: A
authors.lastname: Akinc
authors.affiliation: Alnylam Pharmaceuticals, Cambridge, Massachusetts, United States of America.
authors.forename: Klaus
authors.initials: K
authors.lastname: Charisse
authors.affiliation: Laboratory of Immune Cell Epigenetics and Signaling, Rockefeller University, New York, New York, United States of America.
authors.forename: Alexander
authors.initials: A
authors.lastname: Tarakhovsky
authors.affiliation: Alnylam Pharmaceuticals, Cambridge, Massachusetts, United States of America.
authors.forename: Tatiana I
authors.initials: TI
authors.lastname: Novobrantseva
authors.affiliation: Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
authors.forename: Victor
authors.initials: V
authors.lastname: Koteliansky
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology; D039642:Eukaryotic Initiation Factors / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D041681:NIH 3T3 Cells; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics*
chemical_list: C549050:Ago1 protein, mouse; C526735:Ago2 protein, mouse; C572771:Ago3 protein, mouse; D060565:Argonaute Proteins; D039642:Eukaryotic Initiation Factors; D012333:RNA, Messenger