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Molecular and Cellular Biology, February 1999, p. 1558-1568, Vol. 19, No. 2
Department of Biochemistry, Molecular Biology
and Cell Biology, Northwestern University, Evanston, Illinois
602081;
Department of Cell Biology and
Genetics, Erasmus University School of Medicine, Rotterdam 3000, Holland2; and
Institute of Molecular
and Cell Biology, University of Tartu, Tartu EE2400,
Estonia3
Received 2 September 1998/Returned for modification 8 October
1998/Accepted 26 October 1998
We found previously that neither a 6-kbp promoter fragment nor even
a 120-kbp yeast artificial chromosome (YAC) containing the whole GATA-3
gene was sufficient to recapitulate its full transcription pattern
during embryonic development in transgenic mice. In an attempt to
further identify tissue-specific regulatory elements modulating the
dynamic embryonic pattern of the GATA-3 gene, we have examined the
expression of two much larger (540- and 625-kbp) GATA-3 YACs in
transgenic animals. A lacZ reporter gene was first inserted
into both large GATA-3 YACs. The transgenic YAC patterns were then
compared to those of embryos bearing the identical lacZ
insertion in the chromosomal GATA-3 locus (creating GATA-3/lacZ "knock-ins"). We found that most of the
YAC expression sites and tissues are directly reflective of the
endogenous pattern, and detailed examination of the integrated YAC
transgenes allowed the general localization of a number of very distant
transcriptional regulatory elements (putative central nervous system-,
endocardium-, and urogenital system-specific enhancers). Remarkably,
even the 625-kbp GATA-3 YAC, containing approximately 450 kbp and
150 kbp of 5' and 3' flanking sequences, respectively, does not contain the full transcriptional regulatory potential of the endogenous locus
and is clearly missing regulatory elements that confer tissue-specific expression to GATA-3 in a subset of neural crest-derived cell lineages.
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Localization of Distant Urogenital System-, Central Nervous
System-, and Endocardium-Specific Transcriptional Regulatory
Elements in the GATA-3 Locus
*
Corresponding author. Mailing address: Department of
Biochemistry, Molecular Biology and Cell Biology, Northwestern
University, 2153 N. Campus Dr., Evanston, IL 60208-3500. Phone: (847)
491-5139. Fax: (847) 467-2152. E-mail: d-engel{at}nwu.edu.
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