Partial rescue of GATA-3 by yeast artificial chromosome transgenes.
GATA-3 is essential for murine embryonic development, but elucidating the genetic controls over the complex temporal and tissue-specific transcriptional regulatory pattern of this transcription factor gene has been problematic. Here we report the isolation and characterization of two yeast artificial chromosomes (YACs) bearing the murine GATA-3 gene. Ordered deletions of both YACs show that they define a 1-megabase pair contig spanning the GATA-3 locus. We found that a 120-kb YAC transgene, including 35 kb of 5' as well as 60 kb of 3' flanking sequence, confers normal GATA-3 expression at sites not revealed previously through analysis of plasmid transgenic lines. However, even this 120-kb YAC does not contain sufficient information to recapitulate the complete GATA-3 expression program during embryogenesis. While not complete in its regulatory capacity, the YAC transgene is nonetheless able to complement several homozygous GATA-3 mutant phenotypes and thereby prolong embryonic life.
Lakshmanan
G
G
Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois, 60208, USA.
Lieuw
K H
KH
Grosveld
F
F
Engel
J D
JD
D000818:Animals; D018244:Chromosomes, Artificial, Yeast; D004268:DNA-Binding Proteins / Q000235:genetics*; D050990:GATA3 Transcription Factor; D018507:Gene Expression Regulation, Developmental; D018014:Gene Transfer Techniques; D051379:Mice; D012441:Saccharomyces cerevisiae / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics*
D004268:DNA-Binding Proteins; D050990:GATA3 Transcription Factor; C494748:Gata3 protein, mouse; D015534:Trans-Activators