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Fully modified 2' MOE oligonucleotides redirect polyadenylation
T A Vickers1, J R Wyatt, T Burckin
1Isis Pharmaceuticals, Department of Molecular and Structural Biology, 2280 Faraday Avenue, Carlsbad, CA 92008, USA. tvickers@isisph.com
Nucleic Acids Research
|March 10, 2001
Summary
Antisense oligonucleotides can redirect polyadenylation site selection, leading to increased mRNA stability and altered protein expression. This novel approach enhances, rather than reduces, specific mRNA abundance.
Area of Science:
- Molecular Biology
- Gene Regulation
- RNA Processing
Background:
- Alternative polyadenylation (APA) signals influence mRNA stability by determining the inclusion of destabilization motifs.
- The selection of polyadenylation sites is a critical regulatory mechanism affecting gene expression levels.
Purpose of the Study:
- To investigate the use of antisense oligonucleotides to manipulate polyadenylation site selection.
- To determine if redirecting polyadenylation can alter mRNA stability and protein expression.
Main Methods:
- Utilized modified 2'-O-methoxyethyl/phosphorothioate oligonucleotides targeting the E-selectin 3'-most polyadenylation signal.
- Inhibited polyadenylation at the primary site and induced it at upstream cryptic sites.
Main Results:
- Antisense treatment successfully inhibited polyadenylation at the E-selectin 3'-most site.
- Polyadenylation was redirected to upstream cryptic sites, resulting in shorter transcripts.
- Shorter transcripts contained fewer destabilization sequences, leading to increased mRNA stability.
- Altered protein expression was observed following the manipulation of mRNA stability.
Conclusions:
- Antisense oligonucleotides can effectively redirect polyadenylation site selection.
- This strategy represents a novel method to increase mRNA abundance by enhancing stability.
- Demonstrates the first use of oligonucleotides to increase, rather than decrease, message abundance.