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Journal Article

Single-stranded antisense siRNAs guide target RNA cleavage in RNAi

MPS-Authors
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Urlaub,  H.
Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society;

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Luehrmann,  R.
Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society;

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Tuschl,  T.
Research Group of Combinatorical Biochemistry, MPI for biophysical chemistry, Max Planck Society;

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Citation

Martinez, J., Patkaniowska, A., Urlaub, H., Luehrmann, R., & Tuschl, T. (2002). Single-stranded antisense siRNAs guide target RNA cleavage in RNAi. Cell, 110(5), 563-574. Retrieved from http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6WSN-4C5HD56-5-1&_cdi=7051&_user=38661&_pii=S009286740200908X&_origin=search&_coverDate=09%2F06%2F2002&_sk=998899994&view=c&wchp=dGLbVlb-zSkzV&md5=e60d5b61ed34db921ee5f181faeafa4b&ie=/sdarticle.pdf.


Cite as: https://hdl.handle.net/11858/00-001M-0000-0012-F2FE-F
Abstract
Small interfering RNAs (siRNAs) are the mediators of mRNA degradation in the process of RNA interference (RNAi). Here, we describe a human biochemical system that recapitulates siRNA- mediated target RNA degradation. By using affinity-tagged siRNAs, we demonstrate that a single-stranded siRNA resides in the RNA-induced silencing complex (RISC) together with eIF2C1 and/or eIF2C2 (human GERp95) Argonaute proteins. RISC is rapidly formed in HeLa cell cytoplasmic extract supplemented with 21 nt siRNA duplexes, but also by adding single-stranded antisense RNAs, which range in size between 19 and 29 nucleotides. Single-stranded antisense siRNAs are also effectively silencing genes in HeLa cells, especially when 5'- phosphorylated, and expand the repertoire of RNA reagents suitable for gene targeting.