Antisense RNA inhibition of HPRT synthesis.
The abundant production of antisense hypoxanthine phosphoribosyltransferase (HPRT) RNA in NIH-3T3, COS, or HeLa cells leads to an inhibition of HPRT synthesis. HPRT enzyme levels in cells transfected with mouse HPRT antisense RNA expression vectors are reduced to less than 1% of parental cell activity, resulting in resistance to 6-thioguanine (6TG). The expression of antisense HPRT RNA leads to a marked reduction in the steady-state levels of endogenous HPRT mRNA. Furthermore, we demonstrate that intron-specific antisense RNA, complementary to sequences adjacent to splice donor or acceptor sites of the first intron of the mouse HPRT gene, are effective in depressing endogenous HPRT levels. These studies suggest that antisense RNA can inhibit gene expression in the nucleus, possibly by perturbing nuclear RNA processing.
Stout
J T
JT
Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.
Caskey
C T
CT
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D004351:Drug Resistance / Q000235:genetics; D005091:Exons / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D005839:Gentamicins; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics* / Q000378:metabolism; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010641:Phenotype; D010957:Plasmids; D012323:RNA Processing, Post-Transcriptional; D012326:RNA Splicing / Q000502:physiology; D016372:RNA, Antisense / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D013866:Thioguanine; D014162:Transfection
D005839:Gentamicins; D016372:RNA, Antisense; D012333:RNA, Messenger; C010680:antibiotic G 418; D007041:Hypoxanthine Phosphoribosyltransferase; D013866:Thioguanine