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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)
 
 
 

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.[1]

References

  1. Retroviral-mediated transfer and expression of human beta-globin genes in cultured murine and human erythroid cells. Weber-Benarous, A., Cone, R.D., London, I.M., Mulligan, R.C. J. Biol. Chem. (1988) [Pubmed]
 
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