Generation of Ski-knockdown mice by expressing a long double-strand RNA from an RNA polymerase II promoter.
We have developed a new vector, named pDECAP, to express long double-strand RNA (ds-RNA) from an RNA polymerase II (Pol II) promoter. Because the transcripts from pDECAP lack both the 5'-cap structure and the 3'-poly(A) tail that facilitate ds-RNA export to the cytoplasm, long ds-RNA from pDECAP does not induce the interferon response. Transgenic mice embryos expressing long ds-RNA for the transcriptional corepressor Ski from this vector exhibited phenotypes that were remarkably similar to those of Ski-deficient embryos, including defects of neural tube closure and eye formation. Thus, this vector provides a new tool to efficiently generate tissue-specific knockdown mice for studying gene function in whole animal systems.
Shinagawa
Toshie
T
Laboratory of Molecular Genetics, RIKEN Tsukuba Institute, Tsukuba, Ibaraki 305-0074, Japan.
Ishii
Shunsuke
S
D020121:5' Untranslated Regions / Q000235:genetics; D000015:Abnormalities, Multiple / Q000196:embryology / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003433:Crosses, Genetic; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D004622:Embryo, Mammalian; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D005822:Genetic Vectors; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014162:Transfection
D020121:5' Untranslated Regions; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D012330:RNA, Double-Stranded; D011994:Recombinant Proteins; D012097:Repressor Proteins; C512653:Ski protein, mouse; D008156:Luciferases; D012319:RNA Polymerase II