Abstract
We have examined the effect of the site of integration on the expression of cloned genes introduced into cultured erythroid cells. Smithies et al. [Smithies, O., Gregg, R.G., Boggs, S.S., Koralewski, M.A. & Kucherlapati, R.S. (1985) Nature (London) 317, 230-234] reported the targeted integration of DNA into the human beta-globin locus on chromosome 11 in a mouse erythroleukemia-human cell hybrid. These hybrid cells can undergo erythroid differentiation leading to greatly increased mouse and human beta-globin synthesis. By transfection of these hybrid cells with a plasmid carrying a modified human beta-globin gene and a foreign gene composed of the coding sequence of the bacterial neomycin-resistance gene linked to simian virus 40 transcription signals (SVneo), cells were obtained in which the two genes are integrated at the beta-globin locus on human chromosome 11 or at random sites. When we examined the response of the integrated genes to cell differentiation, we found that the genes inserted at the beta-globin locus were induced during differentiation, whereas randomly positioned copies were not induced. Even the foreign SVneo gene was inducible when it had been integrated at the beta-globin locus. The results show that genes introduced at the beta-globin locus acquire some of the regulatory properties of globin genes during erythroid differentiation.
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More From: Proceedings of the National Academy of Sciences of the United States of America
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