Sensitivity of splice sites to antisense oligonucleotides in vivo.

Sierakowska, H; Sambade, M J; Schümperli, Daniel; Kole, R (1999). Sensitivity of splice sites to antisense oligonucleotides in vivo. RNA - a publication of the RNA Society, 5(3), pp. 369-377. Cold Spring Harbor Laboratory Press

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A series of HeLa cell lines which stably express beta-globin pre-mRNAs carrying point mutations at nt 654, 705, or 745 of intron 2 has been developed. The mutations generate aberrant 5' splice sites and activate a common 3' cryptic splice site upstream leading to aberrantly spliced beta-globin mRNA. Antisense oligonucleotides, which in vivo blocked aberrant splice sites and restored correct splicing of the pre-mRNA, revealed major differences in the sensitivity of these sites to antisense probes. Although the targeted pre-mRNAs differed only by single point mutations, the effective concentrations of the oligonucleotides required for correction of splicing varied up to 750-fold. The differences among the aberrant 5' splice sites affected sensitivity of both the 5' and 3' splice sites; in particular, sensitivity of both splice sites was severely reduced by modification of the aberrant 5' splice sites to the consensus sequence. These results suggest large differences in splicing of very similar pre-mRNAs in vivo. They also indicate that antisense oligonucleotides may provide useful tools for studying the interactions of splicing machinery with pre-mRNA.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Department of Biology > Institute of Cell Biology
08 Faculty of Science > Department of Biology > Institute of Cell Biology > RNA

UniBE Contributor:

Schümperli, Daniel

Subjects:

500 Science > 570 Life sciences; biology
500 Science

ISSN:

1355-8382

Publisher:

Cold Spring Harbor Laboratory Press

Language:

English

Submitter:

Daniel Schümperli

Date Deposited:

14 Sep 2015 09:13

Last Modified:

14 Sep 2015 09:13

PubMed ID:

10094306

URI:

https://boris.unibe.ch/id/eprint/71622

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