Retroviral-mediated transfer and expression of human beta-globin genes in cultured murine and human erythroid cells.
We have cloned human beta-globin DNA sequences from a genomic library prepared from DNA isolated from the human leukemia cell line K562 and have used the retroviral vector pZip-NeoSV(X)1 to introduce a 3.0-kilobase segment encompassing the globin gene into mouse erythroleukemia cells. Whereas the endogenous K562 beta-globin gene is repressed in K562 cells, when introduced into mouse erythroleukemia cells by retroviral-mediated gene transfer, the beta-globin gene from K562 cells was transcribed and induced 5-20-fold after treatment of the cells with dimethyl sulfoxide. The transcripts were correctly initiated, and expression and regulation of the K562 gene were identical to the expression of a normal human beta-globin gene transferred into mouse erythroleukemia cells in the same way. We have also introduced the normal human beta-globin gene into K562 cells using the same retrovirus vector. SP6 analysis of the RNA isolated from the transduced cells showed that the normal beta-globin gene was transcribed at a moderately high level, before or after treatment with hemin. Based on these data, we suggest that the lack of expression of the endogenous beta-globin gene in K562 cells does not result from an alteration in the gene itself and may not result from a lack of factor(s) necessary for beta-globin gene transcription. Retroviral-mediated transfer of the human beta-globin gene may, however, uniquely influence expression of the gene in K562 cells.
Weber-Benarous
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Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142.
Cone
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London
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Mulligan
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D000818:Animals; D002460:Cell Line; D004247:DNA / Q000032:analysis; D004912:Erythrocytes / Q000378:metabolism*; D005914:Globins / Q000235:genetics*; D006427:Hemin / Q000494:pharmacology; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics; D051379:Mice; D012333:RNA, Messenger / Q000096:biosynthesis; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D006427:Hemin; D005914:Globins; D004247:DNA