Selective inhibition of beta-globin RNA transcripts by antisense RNA molecules.
Inhibition of beta-globin gene expression by antisense nucleic acids is a potentially powerful therapeutic strategy for sickle cell disease. To develop clinically relevant beta-globin antisense agents we created nine stable mouse erythroleukemia cell lines expressing unique anti-beta-globin RNA transcripts with different potentials for cross-hybridization with gamma-globin mRNA. We observed variable inhibition of beta-globin expression independent of the hybridization potential of the respective antisense beta-globin RNA transcript. Similarly, inhibition of gamma-globin expression by anti-beta transcripts varied widely in the nine stable cell lines. Three neighboring regions in the beta-globin gene with low RNA folding potentials conferred significantly stronger antisense effect toward beta-globin while sparing the homologous targets in gamma-globin. We have identified for the first time targets in the beta-globin gene for which the homologous regions in gamma-globin are relatively inaccessible to antisense attack. Our findings offer the prospect of using this approach to reduce the proportion of intracellular hemoglobin S. Gene therapy strategies which combine gamma-globin induction along with beta-globin inhibition using antisense vectors may yield more favorable anti-sickling effects longterm.
Pace
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University of Texas at Dallas, Department of Molecular and Cell Biology, PO Box 830688, Mail Station FO 3.1, Richardson, TX 75083, USA. bpace@utdallas.edu
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Ofori-Acquah
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D000818:Animals; D045744:Cell Line, Tumor; D015536:Down-Regulation; D005914:Globins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics; D051379:Mice; D016372:RNA, Antisense / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D014158:Transcription, Genetic / Q000235:genetics*; D014162:Transfection
D016372:RNA, Antisense; D012333:RNA, Messenger; D005914:Globins