RNA interference for the organizer-specific gene Xlim-1 in Xenopus embryos.
Double-stranded RNA (dsRNA) interferes with gene expression in various species, a phenomenon known as RNA interference (RNAi). We show here that RNAi is also effective in modifying gene expression in Xenopus embryos. First, expression of an exogenous luciferase gene as a reporter in embryos was reduced by coinjection with dsRNA corresponding to the luciferase gene. Next, injection of dsRNA for Xlim-1, a homeobox gene suggested to be involved in Spemann organizer functions, reduced the endogenous level of Xlim-1 mRNA and produced embryos with reduced eyes or anterior truncation at high efficiency. In addition, injection of an antisense expression construct of Xlim-1 elicited phenotypes very similar to those of Xlim-1 dsRNA-injected embryos. These results indicate the effectiveness of RNAi for loss of function studies in Xenopus embryos, and the importance of Xlim-1 in head formation.
Laboratory of Molecular Embryology, Department of Biological Sciences, Graduate School of Science, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
H
H
Nakano
S
S
Amemiya
K
K
Shiokawa
M
M
Taira
D000818:Animals; D001757:Blastomeres / Q000187:drug effects / Q000378:metabolism; D016373:DNA, Antisense / Q000494:pharmacology; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000235:genetics*; D017930:Genes, Reporter; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D060850:LIM-Homeodomain Proteins; D008156:Luciferases / Q000235:genetics; D008648:Mesoderm / Q000187:drug effects / Q000473:pathology; D008845:Microinjections; D010641:Phenotype; D016372:RNA, Antisense / Q000494:pharmacology; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors; D029867:Xenopus Proteins; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D016373:DNA, Antisense; D018398:Homeodomain Proteins; D060850:LIM-Homeodomain Proteins; C543964:Lhx1 protein, Xenopus; D016372:RNA, Antisense; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D014157:Transcription Factors; D029867:Xenopus Proteins; D008156:Luciferases; D001616:beta-Galactosidase