An efficient genetic screen in mammalian cultured cells.
Genetic approaches in mammalian cultured cells had limited success because the isolation of mutants and the identification of the mutated genes were often difficult. In the present report, we describe the establishment of a novel genetic screen in Cos-7 cells that allows rapid identification of polypeptides whose overexpression inhibits a certain cellular process. We demonstrate that this approach can be used successfully to isolate partial cDNAs whose overexpression specifically interfered with the clathrin-mediated endocytosis of transferrin.
Schmelzl
Birgit
B
Biochemiezentrum, Heidelberg, Germany.
Geli
M Isabel
MI
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D019556:COS Cells; D002469:Cell Separation; D002478:Cells, Cultured; D002966:Clathrin / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology; D005434:Flow Cytometry; D017930:Genes, Reporter; D005820:Genetic Testing / Q000379:methods*; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011990:Receptors, Transferrin / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D014168:Transferrin / Q000378:metabolism
D002966:Clathrin; D018076:DNA, Complementary; D008164:Luminescent Proteins; D010455:Peptides; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins; D014168:Transferrin; D049452:Green Fluorescent Proteins