Amplification of the human alpha-globin gene enhances its expression.
We investigated the effect of globin gene amplification on alpha-globin gene expression. Chinese hamster ovary cells were transformed with a plasmid containing the human alpha-globin gene linked to a functional dihydrofolate reductase gene, and the transformed cells were selected with increasing concentrations of methotrexate in the culture medium. A cell clone which was resistant to 0.1 mM methotrexate showed a 500-fold amplification of the transformed alpha-globin and DHFR genes. Both genes expressed high levels of their respective mRNAs. The mRNAs were functional and were translated to alpha-globin and DHFR proteins. Thus, one strategy for globin gene therapy is to achieve a high copy number of the transformed globin genes.
D000818:Animals; D002460:Cell Line; D002874:Chromosome Mapping; D006224:Cricetinae; D004274:DNA, Recombinant; D004351:Drug Resistance; D005260:Female; D005784:Gene Amplification; D005786:Gene Expression Regulation; D005914:Globins / Q000235:genetics*; D006801:Humans; D008727:Methotrexate / Q000494:pharmacology; D009693:Nucleic Acid Hybridization; D010053:Ovary / Q000187:drug effects / Q000201:enzymology; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics; D014158:Transcription, Genetic
D004274:DNA, Recombinant; D005914:Globins; D013762:Tetrahydrofolate Dehydrogenase; D008727:Methotrexate