Use of yeast artificial chromosomes (YACs) for studying control of gene expression: correct regulation of the genes of a human beta-globin locus YAC following transfer to mouse erythroleukemia cell lines.
We demonstrate that transfer of a yeast artificial chromosome (YAC) containing 230 kb of the human beta-globin locus into mouse erythroleukemia cells by fusion results in correct developmental regulation of the human beta-like globin genes. Additionally, we show that early after hybrid formation, human embryonic epsilon- and fetal gamma-globin genes are coexpressed with the adult beta gene but that after 10-20 weeks in culture, globin gene expression switches to predominantly adult. Thus, in contrast to shorter gene constructs, the globin genes of the beta-globin locus YAC are regulated like the chromosomal globin genes. These results indicate that transfer of YACs into established cell lines can be used for the analysis of the developmental control of multigenic and developmentally regulated human loci.
Peterson
K R
KR
Division of Medical Genetics, University of Washington, Seattle 98195.
Zitnik
G
G
Huxley
C
C
Lowrey
C H
CH
Gnirke
A
A
Leppig
K A
KA
Papayannopoulou
T
T
Stamatoyannopoulos
G
G
D000818:Animals; D018244:Chromosomes, Artificial, Yeast; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005914:Globins / Q000235:genetics*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics; D015533:Transcriptional Activation
D012333:RNA, Messenger; D005914:Globins