ZFIN ID: ZDB-FIG-200421-35
Adusumilli et al., 2020 - miR-7 Controls the Dopaminergic/Oligodendroglial Fate through Wnt/β-catenin Signaling Regulation. Cells   9(3) Full text @ Cells
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage: Long-pec
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Long-pec

Figure 4

miR-7 gene editing decreases miR-7 expression and increases Wnt signaling in the zebrafish diencephalon. (A) Position of the three Crispr/Cas9 guides used to perform gene editing in the zebrafish miR-7a locus. (B) Agarose gel electrophoretic analysis of the Crispr/Cas9-targeted region in the miR-7a gene, PCR-amplified from genomic DNA of control (ctrl) or guide-injected (g) embryos. Smears indicate Crispr/Cas9-induced gene modification. M: size marker; nc: PCR negative control. (CE) Decreased miR-7 expression is observed in crispants injected with the gU + gI1 Crispr/Cas9 guides, compared with the control (ctrl). Signal quantification (R.I.: relative intensity) is shown in E; (***), p < 0.001, n = 6. (FH) Analysis of diencephalic Wnt signaling (green cells within the white dashed ellipse) in the control (ctrl, F) and guide-injected (gU + gI1, G) Wnt:GFP transgenic embryos (G). Signal quantification is shown in (H). The highest Wnt signal increase (80% more than the control level) is reached with the gU + gI1 guide combination; (*) p < 0.05 and (***) p < 0001; (n.s.), not significant; n = 6. All embryonic heads are at 2 dpf and in dorsal (C,D) or lateral (F,G) view, with anterior to the left. te: telencephalon; di: diencephalon.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP ia4Tg control Long-pec diencephalon IFL
ia4Tg + CRISPR1-mir7a-1 + CRISPR2-mir7a-1 standard conditions Long-pec diencephalon IFL
ia4Tg + CRISPR1-mir7a-1 + CRISPR3-mir7a-1 standard conditions Long-pec diencephalon IFL
ia4Tg + CRISPR3-mir7a-1 standard conditions Long-pec diencephalon IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ia4Tg + CRISPR1-mir7a-1 + CRISPR2-mir7a-1 standard conditions Long-pec diencephalon GFP expression increased amount, abnormal
Long-pec diencephalon Wnt signaling pathway increased occurrence, abnormal
ia4Tg + CRISPR1-mir7a-1 + CRISPR3-mir7a-1 standard conditions Long-pec diencephalon GFP expression increased amount, abnormal
Long-pec diencephalon Wnt signaling pathway increased occurrence, abnormal
ia4Tg + CRISPR3-mir7a-1 standard conditions Long-pec diencephalon GFP expression increased amount, abnormal
Long-pec diencephalon Wnt signaling pathway increased occurrence, abnormal
Acknowledgments:
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