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2008•Nucleic Acids Symposium SeriesOpen access

Tc-DNA modified siRNA

Damian Ittig, Daniel Schümperli, Christian J. Leumann

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Abstract

We investigated the biological activity of siRNAs carrying tc-DNA modifications at various positions in the sense strand. The siRNAs were directed to the coding region of the enhanced green fluorescent protein (EGFP) mRNA. HeLa cells were transfected with the EGFP plasmid and variable concentrations of siRNA duplexes. The antisense effect was quantified on the protein level by fluorescence activated cell sorting (FACS). We found that 3'-end modification as well as modifications at 3-4 further positions either in the 3'- or the 5'-region of the sequence were well tolerated, some of them leading to higher repression of gene expression than wild type RNA.

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We investigated the biological activity of siRNAs carrying tc-DNA modifications at various positions in the sense strand. The siRNAs were directed to the coding region of the enhanced green fluorescent protein (EGFP) mRNA. HeLa cells were transfected with the EGFP plasmid and variable concentrations of siRNA duplexes. The antisense effect was quantified on the protein level by fluorescence activated cell sorting (FACS). We found that 3'-end modification as well as modifications at 3-4 further positions either in the 3'- or the 5'-region of the sequence were well tolerated, some of them leading to higher repression of gene expression than wild type RNA.

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Available abstract

We investigated the biological activity of siRNAs carrying tc-DNA modifications at various positions in the sense strand. The siRNAs were directed to the coding region of the enhanced green fluorescent protein (EGFP) mRNA. HeLa cells were transfected with the EGFP plasmid and variable concentrations of siRNA duplexes. The antisense effect was quantified on the protein level by fluorescence activated cell sorting (FACS). We found that 3'-end modification as well as modifications at 3-4 further positions either in the 3'- or the 5'-region of the sequence were well tolerated, some of them leading to higher repression of gene expression than wild type RNA.

Key concepts: Small interfering RNA, Transfection, Molecular biology, Sense (electronics), HeLa, DNA, Green fluorescent protein, Sense strand

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