Specific gene silencing using small interfering RNAs in fish embryos.
Recently, small interfering RNAs (siRNAs) have been used for gene knockdown in mammalian cultured cells, but their utility in fish has remained unexplored. Here we demonstrate a siRNA-mediated gene silencing technique in rainbow trout embryos. We found that siRNAs effectively suppressed the transient expression of episomally located foreign GFP genes at an early developmental stage and inhibited the expression of GFP genes in stable transgenic trout embryos. Similar gene silencing was observed with an siRNA against the endogenous tyrosinase A gene. siRNAs interfered with the expression of maternally inherited mRNA. siRNAs did not affect non-relevant gene expression and siRNAs with a 4 base mismatch did not affect target gene expression. siRNA gene silencing is therefore highly sequence-specific. Our findings are the first evidence that siRNA-mediated gene silencing is effective in fish. This technique could be a powerful tool for studying gene function during embryonic development in aquacultural fish species, zebrafish, and medaka.
Boonanuntanasarn
Surintorn
S
Department of Aquatic Biosciences, Tokyo University of Fisheries, Konan, Minato, Tokyo 108-8477, Japan.
Yoshizaki
Goro
G
Takeuchi
Toshio
T
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D005399:Fishes / Q000196:embryology / Q000235:genetics*; D020868:Gene Silencing; D014442:Monophenol Monooxygenase / Q000235:genetics; D034741:RNA, Small Interfering / Q000502:physiology*; D019076:Transgenes
D017931:DNA Primers; D034741:RNA, Small Interfering; D014442:Monophenol Monooxygenase