Identification of vegetal RNA-localization elements in Xenopus oocytes.
Localized mRNAs have been identified in a large variety of cell types where they contribute to the establishment of cell asymmetries and can function as cell fate determinants. In Xenopus, RNAs that localize to the vegetal cortex during oogenesis function in early embryonic patterning as well as in the development of primordial germ cells. Based on their temporal and spatial localization patterns, vegetally localizing RNAs are referred to as either early-pathway RNAs which transiently localize in the mitochondrial cloud, or as late-pathway RNAs. Vegetal RNA-localization is driven by cis-acting signal sequences that, in most cases, were found to reside in the 3'-UTRs and which are recognized by trans-acting localization factors. Here we describe the methods of how vegetal RNA-localization elements can be identified by injection of fluorescently-labeled or tagged RNAs.
Claussen
Maike
M
Department of Developmental Biochemistry, Göttingen Center for Molecular Biosciences, Justus-von-Liebig-Weg 11, Göttingen, Germany.
Pieler
Tomas
T
D020413:3' Untranslated Regions; D000818:Animals; D003584:Cytological Techniques; D019286:Endopeptidase K / Q000378:metabolism; D005854:Germ Cells / Q000166:cytology; D007763:Lac Operon; D008845:Microinjections; D008856:Microscopy, Fluorescence / Q000379:methods; D008928:Mitochondria / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D014981:Xenopus / Q000378:metabolism*
D020413:3' Untranslated Regions; D012313:RNA; D012260:Ribonucleases; D019286:Endopeptidase K