Volloch V, Schweitzer B, Rits S
Department of Metabolic Regulation, Boston Biomedical Research Institute, MA 02114.
Biochem Biophys Res Commun. 1991 Sep 30;179(3):1593-9. doi: 10.1016/0006-291x(91)91756-3.
The objective of the experiments described in this paper was to determine the feasibility of inhibition of pre-mRNA splicing by antisense RNA in vitro. Three different types of antisense RNA were utilized: antisense RNA complementary to the spliced RNA molecule; antisense RNA complementary to the unprocessed mRNA precursor molecule; and antisense RNA complementary to the 5' and 3' splice junctions. Whereas antisense RNA complementary to mRNA had little effect on splicing, antisense RNAs complementary to mRNA precursor or to splice junctions strongly inhibited splicing of pre-mRNA molecule. The results obtained indicate that the inhibitory effect is most likely due to hybrid formation between pre-mRNA and antisense RNA molecules and that antisense RNA complementary to the exon portion but not to the intron portion of splice junction exhibit an inhibitory effect. This inhibition can be overcome by bringing together 5' and 3' splice junctions via hybrid formation with antisense RNA complementary to the spliced RNA molecule.
本文所述实验的目的是确定体外反义RNA抑制前体mRNA剪接的可行性。使用了三种不同类型的反义RNA：与剪接后的RNA分子互补的反义RNA；与未加工的mRNA前体分子互补的反义RNA；以及与5'和3'剪接连接点互补的反义RNA。与mRNA互补的反义RNA对剪接几乎没有影响，而与mRNA前体或剪接连接点互补的反义RNA强烈抑制前体mRNA分子的剪接。所得结果表明，抑制作用很可能是由于前体mRNA与反义RNA分子之间形成杂交体，并且与剪接连接点的外显子部分而非内含子部分互补的反义RNA表现出抑制作用。通过与与剪接后的RNA分子互补的反义RNA形成杂交体，将5'和3'剪接连接点聚集在一起，可以克服这种抑制作用。
