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Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
Increased siRNA duplex stability correlates with reduced off-target and elevated on-target effects.
RNA 17, 737-749 (2011)
Argonaute (Ago) proteins form the core of RNA-induced silencing complexes (RISCs) and mediate small RNA-guided gene silencing. In RNAi, short interfering RNAs (siRNAs) guide RISCs to complementary target RNAs, leading to cleavage by the endonuclease Ago2. Noncatalytic Ago proteins, however, contribute to RNAi as well but cannot cleave target RNA and often generate off-target effects. Here we show that synthetic siRNA duplexes interact with all Ago proteins, but a functional RISC rapidly assembles only around Ago2. By stabilizing the siRNA duplex, we show that the noncatalytic Ago proteins Ago1, -3, and -4 can be selectively blocked and do not form functional RISCs. In addition, stabilized siRNAs form an Ago2-RISC more efficiently, leading to increased silencing activity. Our data suggest novel parameters for the design of siRNAs with selective activation of the endonuclease Ago2.
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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
argonaute proteins; RNAi; gene silencing; siRNAs
ISSN (print) / ISBN
1355-8382
e-ISSN
1469-9001
Zeitschrift
RNA
Quellenangaben
Band: 17,
Heft: 4,
Seiten: 737-749
Verlag
Cold Spring Harbor Laboratory Press
Nichtpatentliteratur
Publikationen
Begutachtungsstatus
Peer reviewed
Institut(e)
Institute of Molecular Immunology (IMI)