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Figure 1
Generation of an intestine-specific and inducible microRNA (miR)-21 transgenic zebrafish. (A) Schematic diagram of the stem-loop structure of dre-mir-30e-2. The mature dre-miR-30e sequence (underlined) can be replaced by other mature miRNA sequences. (B) Schematic diagram of stem-loop structure of dre-mir-30e-2-based dre-mir-21. The mature miRNA region has been replaced by mature dre-mir-21. (C) Schematic diagram of the DNA construct used to generate ImiR-21 [Tg(fabp2a:Teton-2A-ZsGreen, TRE:mCherry-miR-21)]. (D) Relative quantification of miR-21 expression using RT-qPCR analysis. ImiR-21#1–5 + doxycycline (Dox) represents five independent transgenic lines. Control: wild-type (WT)—Dox and ImiR-21#1–5—Dox zebrafish. The expression level of miR-21 was normalized to that of U6 in the intestine of the WT—Dox control. (E) Intestine-specific inducible miR-21 expression in ImiR-21#4 at 7 days post-fertilization (dpf). Transgenic larvae were treated with 25 μg/mL Dox from 3 to 7 dpf. Scale bar: 50 μm.