Nucleic Acids Research, 1995, Vol. 23, No. 10 1790-1794
© 1995
MOLECULAR BIOLOGY |
Function of the upstream hypersensitive sites of the chicken ß-globin gene cluster in mice
Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases Building 10, Room 8S–239 1Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health Bethesda, MD 20892-1770, USA
* To whom correspondence should be addressed
Received January 13, 1995. Accepted February 6, 1995.
We have shown previously that the chicken ßA-globln gene, with Its 3' enhancer, is expressed In a copy number-dependent manner In transgenic mice. The expression level was low but Increased {small tilde}6-fold upon inclusion of 11 kb of upstream DNA containing four DNase I hypersensitive sites. To study the effect of the individual upstream hypersensitive sites on transgene expression, we produced lines of mice In which the individual upstream sites were linked to the ßA gene and enhancer. RNA levels were measured in blood from adult animals. With each of these four constructs, the level of transgene RNA per DNA copy varied over a >20-fold range. These data suggest that addition of a hypersensitive site to the ßA-globin/enhancer region abrogates its position independent expression. The average ßA-globin expression per copy In the lines carrying an upstream site was comparable with that in lines without an upstream site. Thus, no single upstream hypersensitive site accounts for the higher level of ßA-globin expression seen in mice containing the complete upstream region. We had shown previously that control of the chicken ß-globin cluster is distributed between at least two regions, the ßA/
enhancer and the upstream region. Our current results suggest that the control mediated by the upstream DNA is itself distributed and is not due to a single hypersensitive site.
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