The human CD34 hematopoietic stem cell antigen promoter and a 3′ enhancer direct hematopoietic expression in tissue culture

Timothy C. Burn, Anne B. Satterthwaite, Daniel G. Tenen

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

The human CD34 hematopoietic stem cell antigen is a highly glycosylated type I membrane protein of unknown function. CD34 is expressed on 1% to 4% of bone marrow cells, including pluripotent stem cells and committed progenitors of each hematopoietic lineage. CD34 has also been shown to be expressed on the small vessel endothelium of a variety of tissues and on a subset of bone marrow stromal cells. We have chosen to use the human CD34 gene as model to examine the transcription factors and cis-elements required for stem cell/progenitor cell-specific gene regulation. We show here that the CD34 gene is transcriptionally regulated in tissue culture cells. Using a luciferase reporter gene, we have isolated and characterized an active CD34 promoter. A CD34-luciferase construct, containing 4.5 kb of 5′ flanking DNA from a CD34 genomic clone, was 30-fold more active in CD34+ tissue culture cells than in HeLa cells. Sequences from the 3′ end of the CD34 gene were shown to have enhancing activity in CD34′ T-lymphoblastic RPMI-8402 cells and not in CD34 U937 cells or in nonhematopoietic HeLa cells. We also show that a cytidine-guanosine island in the 5′ end of the CD34 gene is heavily methylated in two CD34 hematopoietic cell lines and demethylated in two CD34+ cell lines. Analysis of the CD34 promoter should result in the identification of stem cell/progenitor cell-specific transcription factors and should provide a means to direct the expression of heterologous genes in hematopoietic stem cells and progenitors.

Original languageEnglish (US)
Pages (from-to)3051-3059
Number of pages9
JournalBlood
Volume80
Issue number12
StatePublished - Dec 15 1992

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Tissue culture
Hematopoietic Stem Cells
Stem cells
Genes
Antigens
Stem Cells
Luciferases
HeLa Cells
Cells
Transcription Factors
Cell Culture Techniques
Bone
Cell Line
Cytidine
Pluripotent Stem Cells
U937 Cells
Guanosine
Mesenchymal Stromal Cells
Reporter Genes
Islands

ASJC Scopus subject areas

  • Hematology

Cite this

The human CD34 hematopoietic stem cell antigen promoter and a 3′ enhancer direct hematopoietic expression in tissue culture. / Burn, Timothy C.; Satterthwaite, Anne B.; Tenen, Daniel G.

In: Blood, Vol. 80, No. 12, 15.12.1992, p. 3051-3059.

Research output: Contribution to journalArticle

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