Cell-specific RNA interference by peptide-inhibited-peptidase-activated siRNAs


Research Article

J RNAi Gene Silencing (2010), 6(2), 422-430

doi: jrgsxx

Published online: 31 December 2010

Full Text: (html | pdf ~286kb | refs)

Cell-specific RNA interference by peptide-inhibited-peptidase-activated siRNAs

Sandra Koehn †, Hendrik W Schaefer †, Mirko Ludwig †, Natja Haag †, Ulrich S Schubert ‡, Lydia Seyfarth †, Diana Imhof ¥, Udo R Markert †, Tobias G Poehlmann †‡*

† Placenta-Lab, Dept of Obstetrics, Friedrich-Schiller-University Jena, Bachstr 18, 07743 Jena, Germany

‡ Laboratory of Organic and Macromolecular Chemistry, Friedrich-Schiller-University Jena, Humboldstr 10, 07743 Jena, Germany

¥ Institute of Biochemistry and Biophysics, Friedrich-Schiller-University Jena, Hans-Knöll-Str 2, 07745 Jena, Germany

*Correspondence toTobias Poehlmann, Email: tobias.poehlmann@uni-jena.de, Tel: +49 3641 930850, Fax: +49 3641 930852

Received: 18 October 2010, Revised: 12 December 2010, Accepted: 16 December 2010

© Copyright The Authors

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ABSTRACT

The use of chemically-synthesized short interfering RNAs (siRNAs) is the key method of choice to manipulate gene expression in mammalian cell cultures and in vivo. Several previous studies have aimed at inducing cell-specific RNA interference (RNAi) in order to use siRNA molecules as therapeutic reagents. Here, we used peptide-inhibited siRNAs that were activated after cleavage by cell-specific peptidases. We show that siRNAs with bound peptide at the antisense strand could be activated in target cells and were able to induce RNAi in a cell-specific manner. Green Fluorescent Protein (GFP) and Signal Transducer and Activator of Transcription (STAT)-3 gene expression were selectively reduced in a JEG-3 human choriocarcinoma cell line expressing the activating enzyme caspase-4, whereas the effect was absent in HEK cells which lacked the enzyme. In JEG-3 cells, reduction of STAT3 gene expression by conventional and peptide-inhibited siRNA led to a decrease in cell proliferation. This suggests that peptide-inhibited siRNAs provide improved cell specificity and offers new opportunities for their therapeutic use.

KEYWORDS: Caspase-4, peptidase, cell-specific siRNA, RNAi

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