title: RNAi revised--target mRNA-dependent enhancement of gene silencing.
abstract: The discovery of RNA interference (RNAi) gave rise to the development of new nucleic acid-based technologies as powerful investigational tools and potential therapeutics. Mechanistic key details of RNAi in humans need to be deciphered yet, before such approaches take root in biomedicine and molecular therapy. We developed and validated an in silico-based model of siRNA-mediated RNAi in human cells in order to link in vitro-derived pre-steady state kinetic data with a quantitative and time-resolved understanding of RNAi on the cellular level. The observation that product release by Argonaute 2 is accelerated in the presence of an excess of target RNA in vitro inspired us to suggest an associative mechanism for the RNA slicer reaction where incoming target mRNAs actively promote dissociation of cleaved mRNA fragments. This novel associative model is compatible with high multiple turnover rates of RNAi-based gene silencing in living cells and accounts for target mRNA concentration-dependent enhancement of the RNAi machinery.
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Dornseifer
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Willkomm
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Rosel Kretschmer-Kazemi
authors.initials: RK
authors.lastname: Far
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Janine
authors.initials: J
authors.lastname: Liebschwager
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Foteini
authors.initials: F
authors.lastname: Beltsiou
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Frank
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Sandra D
authors.initials: SD
authors.lastname: Laufer
authors.affiliation: Institute for Neuro- and Bioinformatics, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Martinetz
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Georg
authors.initials: G
authors.lastname: Sczakiel
authors.affiliation: Institute for Neuro- and Bioinformatics, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Jens Christian
authors.initials: JC
authors.lastname: Claussen
authors.affiliation: Institute of Molecular Medicine, University of Lübeck, 23538 Lübeck, Germany restle@imm.uni-luebeck.de.
authors.forename: Tobias
authors.initials: T
authors.lastname: Restle
mesh_terms: D060565:Argonaute Proteins / Q000378:metabolism; D003198:Computer Simulation; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008957:Models, Genetic; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering