Vickers T A, Wyatt J R, Burckin T, Bennett C F, Freier S M
Isis Pharmaceuticals, Department of Molecular and Structural Biology, 2280 Faraday Avenue, Carlsbad, CA 92008, USA.
Nucleic Acids Res. 2001 Mar 15;29(6):1293-9. doi: 10.1093/nar/29.6.1293.
Many genes have been described and characterized that have alternative polyadenylation signals at the 3'-end of their pre-mRNAs. Many of these same messages also contain destabilization motifs responsible for rapid degradation of the mRNA. Polyadenylation site selection can thus determine the stability of an mRNA. Fully modified 2'-O:-methoxy ethyl/phosphorothioate oligonucleotides that hybridize to the 3'-most polyadenylation site or signal of E-selectin were able to inhibit polyadenylation at this site and redirect it to one of two upstream cryptic sites. The shorter transcripts produced after antisense treatment have fewer destabilization sequences, increased mRNA stability and altered protein expression. This study demonstrates that antisense oligonucleotides can be successfully employed to redirect polyadenylation. This is the first demonstration of the use of oligonucleotides to increase, rather than decrease, abundance of a message.
许多基因已被描述和表征，其前体mRNA的3'端具有可变聚腺苷酸化信号。许多相同的信使RNA还包含负责mRNA快速降解的不稳定基序。因此，聚腺苷酸化位点的选择可以决定mRNA的稳定性。与E-选择素的最3'端聚腺苷酸化位点或信号杂交的完全修饰的2'-O-甲氧基乙基/硫代磷酸酯寡核苷酸能够抑制该位点的聚腺苷酸化，并将其重定向至两个上游隐蔽位点之一。反义处理后产生的较短转录本具有较少的不稳定序列，增加了mRNA稳定性并改变了蛋白质表达。这项研究表明，反义寡核苷酸可以成功用于重定向聚腺苷酸化。这是首次证明使用寡核苷酸来增加而非减少信使RNA的丰度。
