Improving gene silencing of siRNAs via tricyclo-DNA modification

Ittig, Damian; Luisier, Samuel; Weiler, Jan; Schümperli, Daniel; Leumann, Christian J. (2010). Improving gene silencing of siRNAs via tricyclo-DNA modification. Artificial DNA, PNA & XNA, 1(1), pp. 9-16. Austin, Tex.: Landes Bioscience 10.4161/adna.1.1.11385

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;Small interfering RNAs (siRNAs) can be exploited for the selective silencing of disease-related genes via the RNA interference (RNAi) machinery and therefore raise hope for future therapeutic applications. Especially chemically modified siRNAs are of interest as they are expected to convert lead siRNA sequences into effective drugs. To study the potential of tricyclo-DNA (tc-DNA) in this context we systematically incorporated tc-DNA units at various positions in a siRNA duplex targeted to the EGFP gene that was expressed in HeLa cells. Silencing activity was measured by FACS, mRNA levels were determined by RT-PCR and the biostability of the modifed siRNAs was determined in human serum. We found that modifications in the 3'-overhangs in both the sense and antisense strands were compatible with the RNAi machinery leading to similar activities compared to wild type (wt) siRNA. Additional modifications at the 3'-end, the 5'- end and in the center of the sense (passenger) strand were also well tolerated and did not compromise activity. Extensive modifications of the 3'- and the 5'-end in the antisense (guide) strand, however, abolished RNAi activity. Interestingly, modifications in the center of the duplex on both strands, corresponding to the position of the cleavage site by AGO2, increased efficacy relative to wt by a factor of 4 at the lowest concentrations (2 nM) investigated. In all cases, reduction of EGFP fluorescence was accompanied with a reduction of the EGFP mRNA level. Serum stability analysis further showed that 3'-overhang modifications only moderately increased stability while more extensive substitution by tc-DNA residues significantly enhanced biostability.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry
08 Faculty of Science > Department of Biology > Institute of Cell Biology

UniBE Contributor:

Ittig, Damian; Luisier, Samuel; Schümperli, Daniel and Leumann, Christian

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

1949-095X

Publisher:

Landes Bioscience

Language:

English

Submitter:

Christian Leumann-Reifler

Date Deposited:

04 Oct 2013 14:17

Last Modified:

10 Aug 2015 11:12

Publisher DOI:

10.4161/adna.1.1.11385

BORIS DOI:

10.7892/boris.5183

URI:

https://boris.unibe.ch/id/eprint/5183 (FactScience: 209907)

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