TY - JOUR
T1 - RNAi revised - Target mRNA-dependent enhancement of gene silencing
AU - Dornseifer, Simon
AU - Willkomm, Sarah
AU - Far, Rosel Kretschmer Kazemi
AU - Liebschwager, Janine
AU - Beltsiou, Foteini
AU - Frank, Kirsten
AU - Laufer, Sandra D.
AU - Martinetz, Thomas
AU - Sczakiel, Georg
AU - Claussen, Jens Christian
AU - Restle, Tobias
PY - 2015/1/1
Y1 - 2015/1/1
N2 - 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 highmultiple turnover rates of RNAibased gene silencing in living cells and accounts for target mRNA concentration-dependent enhancement of the RNAi machinery.
AB - 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 highmultiple turnover rates of RNAibased gene silencing in living cells and accounts for target mRNA concentration-dependent enhancement of the RNAi machinery.
UR - http://www.scopus.com/inward/record.url?scp=84975313687&partnerID=8YFLogxK
U2 - 10.1093/nar/gkv1200
DO - 10.1093/nar/gkv1200
M3 - Journal articles
C2 - 26578554
AN - SCOPUS:84975313687
SN - 0305-1048
VL - 43
SP - 10623
EP - 10632
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 22
ER -
