Inducible expression of a neo gene integrated into the human alpha-globin gene cluster.
We replaced the 3' flanking region of the human alpha 1-globin gene that binds in vitro the specific transcription factors GATA-1 and AP1/NF-E2, by a neo marker gene using homologous recombination in a MEL (mouse erythroleukemia line) hybrid cell line harbouring a single human chromosome 16. Using an improved method of the neo-positive and HSV-tk negative selection, one correctly targeted clone was obtained out of 164 clones analyzed. In contrast to non-targeted clones, the expression of teh neo gene in the targeted clone acquired the erythroid differentiation-dependent inducibility normally characteristic of the alpha-globin genes. No difference was observed in the expression of the human zeta, alpha 2, alpha 1, or theta-globin genes before and after induction of differentiation between the targeted clone and parental cells. These results indicate that, at least in the experimental system used, the 3' flanking region of the human alpha 1-globin gene can be replaced by an exogenous non-erythroid gene without affecting the regulation of the globin genes contained in the alpha-globin cluster.
Bernet-Grandaud
A
A
CNRS UMR 106, Université Claude-Bernard, Villeurbanne, France.
Ouazana
R
R
Morlé
F
F
Godet
J
J
D002454:Cell Differentiation; D015672:Erythroid Precursor Cells / Q000166:cytology; D015870:Gene Expression; D018390:Gene Targeting; D005914:Globins / Q000235:genetics*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics / Q000473:pathology; D005810:Multigene Family; D011995:Recombination, Genetic; D014407:Tumor Cells, Cultured
D005914:Globins