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Full Activity From Human  -Globin Locus Control Region Transgenes
Requires 5 HS1, Distal -Globin Promoter, and 3
-Globin Sequences
Peter Pasceri,
Dylan Pannell,
Xiumei Wu, and
James Ellis
From the Developmental Biology Program, Cancer and Blood Program, and
Blood Gene Therapy Program, Hospital for Sick Children, Toronto,
Ontario, Canada, and the Department of Molecular and Medical Genetics,
University of Toronto, Toronto, Ontario, Canada.
The locus control region (LCR) activates high-level human  -globin
transgene expression. LCR cassettes composed of 5 HS2-4 linked to
the 815 bp -globin proximal promoter do not express fully. Here, we
show that LCR (5HS2-4) -globin transgenes that also
contain either 5HS1 or the distal promoter fail to express fully in single- and low-copy transgenic mice. In contrast, full expression is obtained in the presence of both 5HS1 and the
distal promoter. Nine factor binding sites were identified in
5HS1, using in vitro DNaseI footprint and gel retardation
assays, and these include a strong Sp1/Sp3 site, four GATA-1 sites, and
two sites that encompass an ACTAAC motif. LCR (5HS1-4)
-globin transgene constructs with the distal promoter deleted or
replaced by spacer DNA show that specific distal promoter sequences are
required for full expression. An LCR (5HS1-4) transgene
construct with truncated downstream -globin gene sequences indicates
that 3 sequences also play an important role. These results show
that full expression of the -globin gene directed by the LCR
requires 5HS1, the distal -globin promoter, and 3
sequences, and has implications for gene therapy construct design and
models of LCR activation.
Blood, Vol. 92 No. 2 (July 15), 1998:
pp. 653-663
© 1998 by the American Society of Hematology.
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