Use of Yeast Artificial Chromosomes (YACs) for Studying Control of Gene Expression: Correct Regulation of the Genes of a Human β-Globin Locus YAC Following Transfer to Mouse Erythroleukemia Cell Lines

• Published in: Proceedings of the National Academy of Sciences - PNAS 1993-12, Vol.90 (23), p.11207-11211
• Main Authors: Peterson, Kenneth R., Zitnik, Galynn, Huxley, Clare, Lowrey, Christopher H., Gnirke, Andreas, Leppig, Kathleen A., Papayannopoulou, Thalia, Stamatoyannopoulos, George
• Format: Article
• Language: English
• Subjects: Adults; Animals; Biological and medical sciences; Cell lines; Chromosomes, Artificial, Yeast; Cosmids; Erythroid cells; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Regulation; Gene Transfer Techniques; Genes; Genetic loci; Globins - genetics; Humans; Hybridity; L cells; Leukemia, Erythroblastic, Acute - genetics; Mice; Molecular and cellular biology; Molecular genetics; RNA, Messenger - genetics; Somatic cells; Transcriptional Activation
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.23.11207

We demonstrate that transfer of a yeast artificial chromosome (YAC) containing 230 kb of the human β-globin locus into mouse erythroleukemia cells by fusion results in correct developmental regulation of the human β-like globin genes. Additionally, we show that early after hybrid formation, human embryonic ε- and fetal γ-globin genes are coexpressed with the adult β 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 β-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.