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 Blood, 15 April 2005, Vol. 105, No. 8, pp. 3346-3352.
Prepublished online as a Blood First Edition Paper on December 30, 2004; DOI 10.1182/blood-2004-03-0987.

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RED CELLS

Differences of globin transgene expression in stably transfected cell lines and transgenic mice

Qiliang Li, David W. Emery, Hemei Han, Jin Sun, Man Yu, and George Stamatoyannopoulos

From the Department of Medicine, Division of Medical Genetics, University of Washington, Seattle.

Previous studies demonstrated that DNase I hypersensitive site -40 (HS-40) of the {alpha}-globin locus is capable of greatly enhancing expression of a hybrid {beta}/{gamma}-globin transcriptional unit in plasmid-transfected murine erythroleukemia (MEL) cells. However, as reported here, this same {gamma}-globin gene expression cassette was only transcribed at trace amounts in erythroid cells of transgenic mice. This lack of expression was not directly attributable to the {beta}/{gamma}-globin transcriptional unit, since this same unit linked to a composite {beta}-globin locus control region was expressed at high levels in transgenic mice. This lack of expression was also not directly attributable to chromosomal position effects, since addition of chromatin insulators failed to increase the frequency of expression. DNase I hypersensitivity and chromatin immunoprecipitation assays demonstrated that the lack of expression was correlated with a closed chromatin structure. We hypothesize that transgenes undergo dynamic changes in chromatin conformation following chromosomal integration and that the discrepant results reported here can be attributed to the relatively high level of chromatin remodeling that occurs in the transgenic mouse model, coupled with the relative inability of the HS-40 element to maintain an open chromatin state under such conditions. (Blood. 2005;105: 3346-3352)


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L. Lisowski and M. Sadelain
Locus control region elements HS1 and HS4 enhance the therapeutic efficacy of globin gene transfer in -thalassemic mice
Blood, December 15, 2007; 110(13): 4175 - 4178.
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