Post-transcriptional suppression of gene expression in Xenopus embryos by small interfering RNA.
Double-stranded RNA (dsRNA) induces gene-specific silencing in organisms from fungi to animals, a phenomenon known as RNA interference (RNAi). RNAi represents an evolutionarily conserved system to protect against aberrant expression of genes and a powerful tool for gene manipulation. Despite reports that RNAi can be induced in vertebrates, severe sequence-non-specific effects of long dsRNA have been documented in various systems. It has recently been shown in cultured mammalian cells that small interfering RNAs (siRNAs) of 21-23 nt can mediate RNAi but bypass the non-specific response induced by longer dsRNAs. However, the effectiveness of siRNAs has not been demonstrated in living vertebrates. In addition, the mechanism of siRNA suppression of gene expression in vertebrate cells remains to be elucidated. Here we show that synthetic 21 nt siRNAs can specifically inhibit the expression of exogenously introduced as well as endogenous genes in the embryos of Xenopus laevis. siRNAs significantly reduced the steady-state amount of both the mRNA and protein of the cognate gene target. Moreover, co-injection of siRNA with the target RNA transcript specifically suppressed the activity of the latter. Taken together, our findings establish siRNA-mediated post-transcriptional suppression of gene expression in Xenopus embryos.
Zhou
Yuan
Y
Institute of Molecular Biology and Department of Biochemistry, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
Ching
Yick-Pang
YP
Kok
Kin Hang
KH
Kung
Hsiang-fu
HF
Jin
Dong-Yan
DY
D000818:Animals; D004625:Embryo, Nonmammalian / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009843:Oligoribonucleotides / Q000008:administration & dosage*; D012330:RNA, Double-Stranded / Q000008:administration & dosage*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000187:drug effects / Q000378:metabolism; D014158:Transcription, Genetic; D014982:Xenopus laevis
D009843:Oligoribonucleotides; D012330:RNA, Double-Stranded; D011993:Recombinant Fusion Proteins; D008156:Luciferases