Petri, Sebastian and Dueck, Anne and Lehmann, Gerhard and Putz, Nicholas and Ruedel, Sabine and Kremmer, Elisabeth and Meister, Gunter (2011) Increased siRNA duplex stability correlates with reduced off-target and elevated on-target effects. RNA, 17 (4). pp. 737-749. ISSN 1355-8382, 1469-9001
Full text not available from this repository. (Request a copy)Abstract
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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PASSENGER-STRAND; CHEMICAL-MODIFICATION; RNA-INTERFERENCE; GUIDE STRAND; ARGONAUTE; RISC; CLEAVAGE; MICRORNAS; AGO2; MECHANISMS; argonaute proteins; RNAi; gene silencing; siRNAs |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I > Prof. Dr. Gunter Meister |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 23 Jun 2020 08:40 |
| Last Modified: | 23 Jun 2020 08:40 |
| URI: | https://pred.uni-regensburg.de/id/eprint/21083 |
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