Nuclear antisense RNA. An efficient new method to inhibit gene expression.
We describe an efficient new antisense RNA method to inhibit gene expression. Antisense RNAs that are retained in the nucleus bind to target transcripts and appear to lead to the degradation of their targets. Antisense RNAs can be expressed and accumulated specifically in the nucleus if they are not polyadenylated at their 3' ends. In antisense expression vectors we use a cis-acting ribozyme to generate 3'-ends independently of the polyadenylation machinery and thereby inhibit transport of RNA molecules from the nucleus to the cytoplasm. We have evaluated this method in the mouse polyoma virus model system, where nuclear antisense transcripts to the viral early transcription region efficiently reduced the level of viral early-strand RNAs. Nonspecific antisense RNA had no effects on viral gene expression. In comparative studies, nuclear antisense RNAs were significantly more effective in downregulating polyoma virus early RNAs than were conventional antisense molecules, which were processed by polyadenylation.
Liu
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Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
Carmichael
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D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002467:Cell Nucleus / Q000378:metabolism*; D015536:Down-Regulation; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D051379:Mice; D008969:Molecular Sequence Data; D011061:Poly A / Q000378:metabolism; D011120:Polyomavirus / Q000235:genetics* / Q000502:physiology; D012313:RNA / Q000096:biosynthesis; D012323:RNA Processing, Post-Transcriptional; D016372:RNA, Antisense / Q000096:biosynthesis / Q000502:physiology*; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis / Q000502:physiology*
D016372:RNA, Antisense; D016337:RNA, Catalytic; D012367:RNA, Viral; C039919:RNA, recombinant; D011061:Poly A; D012313:RNA