Regulated Expression of Genes Inserted at the Human Chromosomal β -globin Locus by Homologous Recombination

• Published in: Proceedings of the National Academy of Sciences - PNAS 1988-06, Vol.85 (11), p.3845-3849
• Main Authors: Nandi, Asit K., Roginski, Raymond S., Gregg, Ronald G., Smithies, Oliver, Skoultchi, Arthur I.
• Format: Article
• Language: English
• Subjects: 550401 - Genetics- Tracer Techniques; ANIMAL CELLS; ANIMALS; BASIC BIOLOGICAL SCIENCES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; Biological and medical sciences; CELL DIFFERENTIATION; Cell Line; Cell lines; Cells; Cellular differentiation; CHROMOSOMES; DAYS LIVING RADIOISOTOPES; DNA; DNA, Recombinant; Erythroid cells; Erythropoiesis; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Regulation; GENE RECOMBINATION; GENE REGULATION; Genes; Genetic loci; GLOBIN; Globins - genetics; HETEROCHROMOSOMES; Humans; Hybrid cells; HYBRIDIZATION; ISOTOPES; LIGHT NUCLEI; MAMMALS; MAN; MESSENGER-RNA; MICE; Molecular and cellular biology; Molecular genetics; NUCLEI; NUCLEIC ACIDS; ODD-ODD NUCLEI; ORGANIC COMPOUNDS; PHOSPHORUS 32; PHOSPHORUS ISOTOPES; Plasmids; PRIMATES; RADIOISOTOPES; Recombination, Genetic; RNA; RNA, Messenger - genetics; RODENTS; TRANSCRIPTION; Transcription, Genetic; TUMOR CELLS; VERTEBRATES
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.85.11.3845

We have examined the effect of the site of integration on the expression of cloned genes introduced into cultured erythroid cells. Smithies et al. [Smithies, O., Gregg, R. G., Boggs, S. S., Koralewski, M. A. & Kucherlapati, R. S. (1985) Nature (London) 317, 230-234] reported the targeted integration of DNA into the human β -globin locus on chromosome 11 in a mouse erythroleukemia-human cell hybrid. These hybrid cells can undergo erythroid differentiation leading to greatly increased mouse and human β -globin synthesis. By transfection of these hybrid cells with a plasmid carrying a modified human β -globin gene and a foreign gene composed of the coding sequence of the bacterial neomycin-resistance gene linked to simian virus 40 transcription signals (SVneo), cells were obtained in which the two genes are integrated at the β -globin locus on human chromosome 11 or at random sites. When we examined the response of the integrated genes to cell differentiation, we found that the genes inserted at the β -globin locus were induced during differentiation, whereas randomly positioned copies were not induced. Even the foreign SVneo gene was inducible when it had been integrated at the β -globin locus. The results show that genes introduced at the β -globin locus acquire some of the regulatory properties of globin genes during erythroid differentiation.