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Prepublished online as a Blood First Edition Paper on August 22, 2002; DOI 10.1182/blood-2002-03-0788.
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Blood, 15 December 2002, Vol. 100, No. 13, pp. 4420-4426
HEMATOPOIESIS
Distal elements are critical for human CD34 expression in
vivo
Yutaka Okuno,
Claudia S. Huettner,
Hanna S. Radomska,
Victoria Petkova,
Hiromi Iwasaki,
Koichi Akashi, and
Daniel G. Tenen
From the Hematology/Oncology Division, Harvard
Institutes of Medicine; and the Department of Cancer Immunology and
AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston,
MA.
The elements regulating gene expression in hematopoietic stem cells
are still poorly understood. We previously reported that a 141-kilobase
(kb) human CD34 transgene confers properly regulated human CD34
expression in transgenic mice. A construct with only the human CD34
promoter and 3' enhancer region is not sufficient, suggesting that
critical distal elements are necessary for expression of the human CD34
gene. To further localize such elements, we analyzed deletion
constructs of the human CD34 gene and evaluated their function in
transgenic mice. Constructs harboring as little as 18 kb of 5' and 26 kb of 3' human CD34 flanking sequence conferred human expression in
tissues of transgenic mice with a pattern similar to that of the 141-kb
human transgene. In contrast, a construct harboring 10 kb of 5' and 17 kb of 3' human CD34 flanking sequence gave no expression. These data
demonstrate that regions between 10 to 18 kb upstream and/or 17 to 26 kb downstream of the human CD34 gene contain critical elements for
human CD34 expression in vivo. Further functional analysis
of these regions in transgenic mice will be crucial for understanding
CD34 gene expression in hematopoietic stem and progenitor cells.

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