Nuclear antisense effects in cyclophilin A pre-mRNA splicing by oligonucleotides: A comparison of tricyclo-DNA with LNA

Ittig, D.; Liu, Shi-Xia; Renneberg, Dorte; Schümperli, Daniel; Leumann, Christian (2004). Nuclear antisense effects in cyclophilin A pre-mRNA splicing by oligonucleotides: A comparison of tricyclo-DNA with LNA. Nucleic acids research, 32(1), pp. 346-353. Oxford University Press 10.1093/nar/gkh187

[img]
Preview
Text
346.pdf - Published Version
Available under License Creative Commons: Attribution-Noncommercial (CC-BY-NC).

Download (243kB) | Preview
[img]
Preview
Text
gkh187.pdf - Other
Available under License Publisher holds Copyright.

Download (243kB) | Preview

The nuclear antisense properties of a series of tricyclo(tc)-DNA oligonucleotide 9-15mers, targeted against the 3' and 5' splice sites of exon 4 of cyclophilin A (CyPA) pre-mRNA, were evaluated in HeLa cells and compared with those of corresponding LNA-oligonucleotides. While the 9mers showed no significant antisense effect, the 11-15mers induced exon 4 skipping and exon 3+4 double skipping to about an equal extent upon lipofectamine mediated transfection in a sequence and dose dependent manner, as revealed by a RT-PCR assay. The antisense efficacy of the tc-oligonucleotides was found to be superior to that of the LNA-oligonucleotides in all cases by a factor of at least 4-5. A tc-oligonucleotide 15mer completely abolished CyPA mRNA production at 0.2‘M concentration. The antisense effect was confirmed by western blot analysis which revealed a reduction of CyPA protein to 13% of its normal level. Fluorescence microscopic investigations with a fluorescein labeled tc-15mer revealed a strong propensity for homogeneous nuclear localization of this backbone type after lipofectamine mediated transfection, while the corresponding lna 15mer showed a less clear cellular distribution pattern. Transfection without lipid carrier showed no significant internalization of both tc- and LNA-oligonucleotides. The obtained results confirm the power of tricyclo-DNA for nuclear antisense applications. Morover, CyPA may become an interesting therapeutic target due to its important role in the early steps of the viral replication of HIV-1.

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:

Liu, Shi-Xia; Renneberg, Dorte; Schümperli, Daniel and Leumann, Christian

Subjects:

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

ISSN:

0305-1048

Publisher:

Oxford University Press

Language:

English

Submitter:

Christian Leumann-Reifler

Date Deposited:

16 Jun 2014 11:08

Last Modified:

27 Apr 2018 08:59

Publisher DOI:

10.1093/nar/gkh187

Uncontrolled Keywords:

analysis Antisense backbone Cells cellular Cyclophilin A distribution double Fluorescein fluorescence HeLa Hela Cells Hiv-1 LNA oligonucleotide Oligonucleotides properties Protein reduction REPLICATION site Transfection tricyclo-DNA

BORIS DOI:

10.7892/boris.53215

URI:

https://boris.unibe.ch/id/eprint/53215

Actions (login required)

Edit item Edit item
Provide Feedback