TY - JOUR
T1 - Multiple elements in human β-globin locus control region 5′ HS 2 are involved in enhancer activity and position independent, transgene expression
AU - Caterina, John J.
AU - Ciavatta, Dominic J.
AU - Donze, David
AU - Behringer, Richard R.
AU - Townes, Tim M.
N1 - Funding Information:
We thank the entire staff of the UAB Transgenic Animal Facility for producing some of the animals described in this study. This work was supported in part by grants HL35559 and HL43508 from the National Institutes of Health. John Caterina is a graduate fellow supported by National Institutes of Health grant 5 T32 CA09467. David Donze is a graduate fellow supported by National Institutes of Health grant T32 GM08111.
PY - 1994/3/25
Y1 - 1994/3/25
N2 - The human β-globin Locus Control Region (LCR) has two important activities. First, the LCR opens a 200 kb chromosomal domain containing the human ε-, γ- and β-giobin genes and, secondly, these sequences function as a powerful enhancer of ε-, γ- and β-globin gene expression. Erythrold-specific, DNase I hypersensitive sites (HS) mark sequences that are critical for LCR activity. Previous experiments demonstrated that a 1.9 kb fragment containing the 5′ HS 2 site confers position-independent expression in transgenic mice and enhances human β-giobin gene expression 100-fold. Further analysis of this region demonstrates that multiple sequences are required for maximal enhancer activity; deletion of SP1, NF-E2, GATA-1 or USF binding sites significantly decrease β-globin gene expression. In contrast, no single site is required for position- independent transgene expression; all mice with site- specific mutations in 5′ HS 2 express human β-globin mRNA regardless of the site of transgene integration. Apparently, multiple combinations of protein binding sites in 5′ HS 2 are sufficient to prevent chromosomal position effects that inhibit transgene expression.
AB - The human β-globin Locus Control Region (LCR) has two important activities. First, the LCR opens a 200 kb chromosomal domain containing the human ε-, γ- and β-giobin genes and, secondly, these sequences function as a powerful enhancer of ε-, γ- and β-globin gene expression. Erythrold-specific, DNase I hypersensitive sites (HS) mark sequences that are critical for LCR activity. Previous experiments demonstrated that a 1.9 kb fragment containing the 5′ HS 2 site confers position-independent expression in transgenic mice and enhances human β-giobin gene expression 100-fold. Further analysis of this region demonstrates that multiple sequences are required for maximal enhancer activity; deletion of SP1, NF-E2, GATA-1 or USF binding sites significantly decrease β-globin gene expression. In contrast, no single site is required for position- independent transgene expression; all mice with site- specific mutations in 5′ HS 2 express human β-globin mRNA regardless of the site of transgene integration. Apparently, multiple combinations of protein binding sites in 5′ HS 2 are sufficient to prevent chromosomal position effects that inhibit transgene expression.
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U2 - 10.1093/nar/22.6.1006
DO - 10.1093/nar/22.6.1006
M3 - Article
C2 - 8152905
AN - SCOPUS:0028357241
SN - 0305-1048
VL - 22
SP - 1006
EP - 1011
JO - Nucleic acids research
JF - Nucleic acids research
IS - 6
ER -