Intracellular expression of anti-p21ras single chain Fv fragments inhibits meiotic maturation of xenopus oocytes.
The recombinant variable regions of the monoclonal antibody Y13-259, directed against the p21ras protein, have been engineered for expression as intracellular single chain Fv fragments. The activity of the plasmid was confirmed by in vitro and in vivo translation of mRNA showing that the intracellularly expressed single chain fragments are stably and efficiently expressed as cytosolic proteins. The expression of the anti-p21ras single chain antibodies in the cytoplasm of Xenopus laevis oocytes leads to the inhibition of the insulin-induced meiotic maturation. This finding represents the first successful application of the strategy of intracellular antibodies to block a complex biological process in the cytosol of vertebrate cells.
Biocca
S
S
Institute of Neurobiology, CNR, Roma, Italy.
Pierandrei-Amaldi
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P
Cattaneo
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000378:metabolism*; D005260:Female; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology; D008540:Meiosis / Q000187:drug effects / Q000502:physiology*; D008845:Microinjections; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D014176:Protein Biosynthesis; D016283:Proto-Oncogene Proteins p21(ras) / Q000276:immunology*; D012333:RNA, Messenger / Q000008:administration & dosage / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D015183:Restriction Mapping; D012156:Reticulocytes / Q000378:metabolism; D014158:Transcription, Genetic; D014714:Vertebrates; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D007328:Insulin; D012333:RNA, Messenger; D011994:Recombinant Proteins; D016283:Proto-Oncogene Proteins p21(ras)