Renneberg, Dorte; Bouliong, Emilie; Reber, Ulrich; Schümperli, Daniel; Leumann, Christian J. (2002). Antisense properties of tricyclo-DNA. Nucleic acids research, 30(13), pp. 2751-2757. Oxford University Press 10.1093/nar/gkf412
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Tricyclo (tc)-DNA belongs to the class of conformationally constrained DNA analogs that show enhanced binding properties to DNA and RNA. We prepared tc-oligonucleotides up to 17 nt in length, and evaluated their binding efficiency and selectivity towards complementary RNA, their biological stability in serum, their RNase H inducing potential and their antisense activity in a cellular assay. Relative to RNA or 2'-O-Me-phosphorothioate (PS)-RNA, fully modified tc-oligodeoxynucleotides, 10-17 nt in length, show enhanced selectivity and enhanced thermal stability by approximately 1 degrees C/modification in binding to RNA targets. Tricyclodeoxyoligonucleotides are completely stable in heat-deactivated fetal calf serum at 37 degree C. Moreover, tc-DNA-RNA duplexes are not substrates for RNase H. To test for antisense effects in vivo, we used HeLa cell lines stably expressing the human beta-globin gene with two different point mutations in the second intron. These mutations lead to the inclusion of an aberrant exon in beta-globin mRNA. Lipofectamine-mediated delivery of a 17mer tc-oligodeoxynucleotide complementary to the 3'-cryptic splice site results in correction of aberrant splicing already at nanomolar concentrations with up to 100-fold enhanced efficiency relative to a 2'-O-Me-PS-RNA oligonucleotide of the same length and sequence. In contrast to 2'-O-Me-PS-RNA, tc-DNA shows antisense activity even in the absence of lipofectamine, albeit only at much higher oligonucleotide concentrations.
Item Type: |
Journal Article (Original Article) |
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Division/Institute: |
08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP) 08 Faculty of Science > Department of Biology > Institute of Cell Biology |
UniBE Contributor: |
Renneberg, Dorte, Schümperli, Daniel, Leumann, Christian |
Subjects: |
500 Science > 570 Life sciences; biology 500 Science > 540 Chemistry |
ISSN: |
0305-1048 |
Publisher: |
Oxford University Press |
Language: |
English |
Submitter: |
Daniel Schümperli |
Date Deposited: |
12 May 2015 08:48 |
Last Modified: |
05 Dec 2022 14:47 |
Publisher DOI: |
10.1093/nar/gkf412 |
PubMed ID: |
12087157 |
BORIS DOI: |
10.7892/boris.68853 |
URI: |
https://boris.unibe.ch/id/eprint/68853 |
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