Molecular Biology of the Cell · 1999 · 623 citations · 59 references
Life SciencesCell BiologyGeneticsEndocytosisGene ExpressionMembrane TraffickingClathrin-mediated EndocytosisLate EndosomesReceptor-mediated EndocytosisRna InterferenceSecretory PathwayVesicle TraffickingC. Elegans Developmental ModelsEndocytic InternalizationEndocytic Trafficking Of ReceptorsEndocytosis PathwaysGreen Fluorescent Protein ReportersLow-density Lipoprotein Receptor FamilyLow-density Lipoprotein Receptor GeneticsRna Interference Constructs
The C. elegans oocyte provides a powerful system for genetic studies of receptor‑mediated endocytosis. The study aims to use YP170::GFP transgenic strains to monitor yolk endocytosis in vivo. YP170::GFP reporter strains were employed to assay endocytic protein functions via RNAi and to screen for rme mutants.
The Caenorhabditis elegans oocyte is a highly amenable system for forward and reverse genetic analysis of receptor-mediated endocytosis. We describe the use of transgenic strains expressing a vitellogenin::green fluorescent protein (YP170::GFP) fusion to monitor yolk endocytosis by theC. elegans oocyte in vivo. This YP170::GFP reporter was used to assay the functions of C. eleganspredicted proteins homologous to vertebrate endocytosis factors using RNA-mediated interference. We show that the basic components and pathways of endocytic trafficking are conserved between C. elegans and vertebrates, and that this system can be used to test the endocytic functions of any new gene. We also used the YP170::GFP assay to identify rme(receptor-mediated endocytosis) mutants. We describe a new member of the low-density lipoprotein receptor superfamily, RME-2, identified in our screens for endocytosis defective mutants. We show that RME-2 is the C. elegans yolk receptor.
59
THE GENETICS OF <i>CAENORHABDITIS ELEGANS</i>
Sydney Brenner · Genetics · 1974 · 15.5K citations · Full text
Life Sciences, Biology, Genetics +14
Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans
Andrew Fire, Siqun Xu, Mary K. Montgomery et al. · Nature · 1998 · 15.3K citations · Full text
Green Fluorescent Protein as a Marker for Gene Expression
Martin Chalfie, Yuan Tu, Ghia Euskirchen et al. · Science · 1994 · 6.8K citations
A new method for predicting signal sequence cleavage sites
Gunnar von Heijne · Nucleic Acids Research · 1986 · 4.6K citations · Full text