Targeted inactivation of the insulin receptor gene in mouse 3T3-L1 fibroblasts via homologous recombination.
To study the role of the insulin receptor in determining adipocyte differentiation of the mouse cell line 3T3-L1, we have introduced a mutation that inactivates the insulin receptor gene by homologous recombination. In two independent clones, inactivation of one allele of the insulin receptor gene was associated with a 50-70% reduction in the number of insulin receptors. In addition, both clones were markedly impaired in their ability to differentiate into adipocytes. The defect in adipocyte-specific differentiation was corrected by expression of transfected human insulin receptor cDNA. These data suggest that the insulin receptor may play an important role in promoting differentiation of 3T3-L1 cells into adipocytes in vitro.
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Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.
Taylor
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D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002454:Cell Differentiation; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005786:Gene Expression Regulation; D015321:Gene Rearrangement; D005822:Genetic Vectors; D006801:Humans; D007739:L Cells / Q000166:cytology / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000235:genetics; D011972:Receptor, Insulin / Q000235:genetics*; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014162:Transfection
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D004247:DNA; D011972:Receptor, Insulin