Oates, A. C.Bruce, A. E. E.Ho, R. K.2017-05-302017-05-302017-05-30200010.1006/dbio.2000.97612-s2.0-0034254815https://infoscience.epfl.ch/handle/20.500.14299/137800We have investigated the ability of double-stranded RNA (dsRNA) to inhibit gene expression in a vertebrate, the zebrafish, Danio rerio. Injection of dsRNA corresponding to the T-box gene tbx16/spadetail (spt) into early wild-type embryos caused a rapid and dramatic loss of tbx16/spt mRNA in the blastula, mRNAs from the papc, tbx6, and gata1 genes, which depend on tbx16/spt function for their expression, were reduced, apparently mimicking the spt mutant phenotype. However, mRNAs from a number of genes that are unaffected by the spt mutation, such as β catenin, stat3, and no tail, were also lost, indicating that the 'interference' effect of tbx16/spt dsRNA was not restricted to the endogenous tbx16/spt mRNA. We compared the effects of injecting dsRNA from the zebrafish tbx16/spadetail, nieuwkoid/bozozok, and Brachyury/no tail genes with dsRNA from the bacterial lacZ gene. In each case the embryos displayed a variable syndrome of abnormalities at 12 and 24 h postfertilization. In blind studies, we could not distinguish between the effects of the various dsRNAs. Consistent with a common effect of dsRNA, regardless of sequence, injection of dsRNA from the lacZ gene was likewise effective in strongly reducing tbx16/spt and β catenin mRNA in the blastula. These findings indicate that, despite published reports, the current methodology of double-stranded RNA interference is not a practical technique for investigating zygotic gene function during early zebrafish development. (C) 2000 Academic Press.animal cellAnimalsArticlecell deathDouble-Strandeddouble stranded RNAembryoembryo developmentgene expression regulationgene functiongene lossgene silencingin situ hybridizationMessengermessenger RNAMicroinjectionsnonhumanNonmammalianphenotypepriority journalRNARNA analysisRNA sequenceT-Box Domain Proteinszebra fishzebrafishZebrafish Proteinstext::journal::journal article::research article