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• W2080555544 abstract "Short interfering RNAs (siRNAs) guide mRNA cleavage during RNA interference (RNAi) [1Elbashir S.M. Lendeckel W. Tuschl T. RNA interference is mediated by 21- and 22-nucleotide RNAs.Genes Dev. 2001; 15: 188-200Crossref PubMed Scopus (2704) Google Scholar, 2Elbashir S.M. Martinez J. Patkaniowska A. Lendeckel W. Tuschl T. Functional anatomy of siRNAs for mediating efficient RNAi in Drosophila melanogaster embryo lysate.EMBO J. 2001; 20: 6877-6888Crossref PubMed Scopus (1210) Google Scholar, 3Schwarz D. Hutvagner G. Haley G. Zamore P.D. siRNAs function as guides, not primers, in the RNAi pathway in Drosophila and human cells.Mol. Cell. 2002; 10: 537-548Abstract Full Text Full Text PDF PubMed Scopus (385) Google Scholar]. Only one siRNA strand assembles into the RNA-induced silencing complex (RISC) [4Martinez J. Patkaniowska A. Urlaub H. Luhrmann R. Tuschl T. Single-stranded antisense siRNAs guide target RNA cleavage in RNAi.Cell. 2002; 110: 563-574Abstract Full Text Full Text PDF PubMed Scopus (1188) Google Scholar], with preference given to the strand whose 5′ terminus has lower base-pairing stability [5Schwarz D.S. Hutvagner G. Du T. Xu Z. Aronin N. Zamore P.D. Asymmetry in the assembly of the RNAi enzyme complex.Cell. 2003; 115: 199-208Abstract Full Text Full Text PDF PubMed Scopus (2198) Google Scholar, 6Khvorova A. Reynolds A. Jayasena S.D. Functional siRNAs and miRNAs exhibit strand bias.Cell. 2003; 115: 209-216Abstract Full Text Full Text PDF PubMed Scopus (2021) Google Scholar]. In Drosophila, Dcr-2/R2D2 processes siRNAs from longer double-stranded RNAs (dsRNAs) [7Liu Q. Rand T.A. Kalidas S. Du F. Kim H.-E. Smith D.P. Wang X. R2D2, a bridge between the initiation and effector steps of the Drosophila RNAi pathway.Science. 2003; 301: 1921-1925Crossref PubMed Scopus (560) Google Scholar, 8Lee Y.S. Nakahara K. Pham J.W. Kim K. He Z. Sontheimer E.J. Carthew R.W. Distinct roles for Drosophila Dicer-1 and Dicer-2 in the siRNA/miRNA silencing pathways.Cell. 2004; 117: 69-81Abstract Full Text Full Text PDF PubMed Scopus (1009) Google Scholar] and also nucleates RISC assembly [7Liu Q. Rand T.A. Kalidas S. Du F. Kim H.-E. Smith D.P. Wang X. R2D2, a bridge between the initiation and effector steps of the Drosophila RNAi pathway.Science. 2003; 301: 1921-1925Crossref PubMed Scopus (560) Google Scholar, 9Pham J.W. Pellino J.L. Lee Y.S. Carthew R.W. Sontheimer E.J. A Dicer-2-dependent 80S complex cleaves targeted mRNAs during RNAi in Drosophila.Cell. 2004; 117: 83-94Abstract Full Text Full Text PDF PubMed Scopus (349) Google Scholar, 10Tomari Y. Du T. Haley B. Schwarz D.S. Bennett R. Cook H.A. Koppetsch B.S. Theurkauf W.E. Zamore P.D. RISC assembly defects in the Drosophila RNAi mutant armitage.Cell. 2004; 116: 831-841Abstract Full Text Full Text PDF PubMed Scopus (325) Google Scholar], suggesting that nascent siRNAs could remain bound to Dcr-2/R2D2. In vitro, Dcr-2/R2D2 senses base-pairing asymmetry of synthetic siRNAs and dictates strand selection by asymmetric binding to the duplex ends [11Tomari Y. Matranga C. Haley B. Martinez N. Zamore P.D. A protein sensor for siRNA asymmetry.Science. 2004; 306: 1377-1380Crossref PubMed Scopus (469) Google Scholar]. During dsRNA processing, Dicer (Dcr) liberates siRNAs from dsRNA ends [1Elbashir S.M. Lendeckel W. Tuschl T. RNA interference is mediated by 21- and 22-nucleotide RNAs.Genes Dev. 2001; 15: 188-200Crossref PubMed Scopus (2704) Google Scholar, 12Zhang H. Kolb F.A. Brondani V. Billy E. Filipowicz W. Human Dicer preferentially cleaves dsRNAs at their termini without a requirement for ATP.EMBO J. 2002; 21: 5875-5885Crossref PubMed Scopus (480) Google Scholar] in a manner dictated by asymmetric enzyme-substrate interactions [13Zhang H. Kolb F.A. Jaskiewicz L. Westhof E. Filipowicz W. Single processing center models for human Dicer and bacterial RNase III.Cell. 2004; 118: 57-68Abstract Full Text Full Text PDF PubMed Scopus (713) Google Scholar]. Because Dcr-2/R2D2 is unlikely to sense base-pairing asymmetry of an siRNA that is embedded within a precursor, it is not clear whether processed siRNAs strictly follow the thermodynamic asymmetry rules [5Schwarz D.S. Hutvagner G. Du T. Xu Z. Aronin N. Zamore P.D. Asymmetry in the assembly of the RNAi enzyme complex.Cell. 2003; 115: 199-208Abstract Full Text Full Text PDF PubMed Scopus (2198) Google Scholar, 6Khvorova A. Reynolds A. Jayasena S.D. Functional siRNAs and miRNAs exhibit strand bias.Cell. 2003; 115: 209-216Abstract Full Text Full Text PDF PubMed Scopus (2021) Google Scholar] or whether processing polarity can affect strand selection [1Elbashir S.M. Lendeckel W. Tuschl T. RNA interference is mediated by 21- and 22-nucleotide RNAs.Genes Dev. 2001; 15: 188-200Crossref PubMed Scopus (2704) Google Scholar]. We use a Drosophila in vitro system in which defined siRNAs with known asymmetry can be generated from longer dsRNA precursors. These dsRNAs permit processing specifically from either the 5′ or the 3′ end of the thermodynamically favored strand of the incipient siRNA. Combined dsRNA-processing/mRNA-cleavage assays indicate that siRNA strand selection is independent of dsRNA processing polarity during Drosophila RISC assembly in vitro." @default.
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• W2080555544 date "2006-03-01" @default.
• W2080555544 modified "2023-10-16" @default.
• W2080555544 title "Short Interfering RNA Strand Selection Is Independent of dsRNA Processing Polarity during RNAi in Drosophila" @default.
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• W2080555544 doi "https://doi.org/10.1016/j.cub.2006.01.061" @default.
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