ZFIN Image: Giacomotto et al., 2015, Fig. 3

Image

Figure Caption

Fig. 3 Evaluation of the effect of target sequence/position on knockdown efficiency in 52 hpf zebrafish embryos.

(a) Schematic representation of mCherry mRNA produced in the βactin:mCherry:sUTR zebrafish line, as well as in artificial miRNA targets. (b–p) Embryos with ubiquitous mCherry expression were injected with transposase and 30pg of tol2-flanked miRNA-expressing constructs (mmu-miR155 backbone), which were designed to produce artificial miRNA targeting different sites of the mCherry mRNA. A minimum of 10 larvae per condition were analysed in order to estimate knockdown efficiency. (q–u) Homozygous βactin:mCherry:sUTR embryos were injected with corresponding RNA (150pg), synthesized via linearized pME-RNAi clones, and raw red fluorescence was quantified, normalized and presented as percentage. The means of 10 embryos±s.d. Different from control at *P=0.05, **P=0.000001. Scale bar, 50µm.

Acknowledgments

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