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A 36-Base-Pair Core Sequence of Locus Control Region Enhances Retrovirally Transferred Human β-Globin Gene Expression

Judy C. Chang, Depei Liu and Yuet Wai Kan
Proceedings of the National Academy of Sciences of the United States of America
Vol. 89, No. 7 (Apr. 1, 1992), pp. 3107-3110
Stable URL: http://www.jstor.org/stable/2359043
Page Count: 4
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Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
A 36-Base-Pair Core Sequence of Locus Control Region Enhances Retrovirally Transferred Human β-Globin Gene Expression
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Abstract

The locus control region of the human β-globin cluster consists of four major DNase I hypersensitive sites (HS). When linked to globin genes, the locus control region confers a high level of erythroid-specific expression of these genes in transgenic mice or transfected erythroid cell lines. We have examined the effect of one of these sites, HS2, on human β-globin gene expression in a murine erythroleukemia cell line (MEL) after retrovirus-mediated gene transfer. We incorporated a 732- or 412-base-pair (bp) segment of HS2 in the retroviral construct carrying the human β-globin gene. These fragments rendered the viruses unstable as the human β-globin gene was rearranged or deleted in all the packaging cell lines examined. On the other hand, when a 36-bp fragment containing the NFE-2/AP-1 binding consensus in this region was inserted into the retroviral construct, we recovered 6 stable packaging cell lines of 12 examined, similar in percentage to the construct with the β-globin gene alone. The virus titers of the packaging cell lines from these two constructs were similar. We infected MEL cells with viruses produced from three packaging cell lines of each of the two constructs and measured the ratio of human β-globin to mouse α-globin mRNA after hexamethylenebisacetamide induction. The overall level of expression increased 2-fold from 6.0% to 12.7% with the addition of this 36-bp enhancer.

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