Shanghai Model Organisms Center,Inc
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SMOC Zebrafish Service Platform

    World-class facilities and equipments
    Achievement of accreditation from AAALAC International
    Genetically modified zebrafish models (Morpholino Knock down, CRISPR/Cas9 Knock out and Transgenic zebrafish)
    Various services:  gene function analysis, translational medicine research, drug screening,  multiple pharmacodynamics index and toxicity evaluation…
 
Case Study
 Zebrafish gene X morphants exhibit a Hedgehog loss-of-function phenotype
 
Fig.1. gene X knock down exhibit a Hedgehog loss-of-function phenotype. (A-B) Gross morphology at 52 hours post fertilization (hpf). (C-D) Somite morphology in wild-type (WT) (C) or in embryos injected with gene X-MO (D). Compared with WT, knock down gene X mimics misregulation of the Hedgehog pathway, displaying characteristic U-shaped somites (red arrow) 、 curved body axis (blue dotted line) and pericardial oedema (black arrow) (See Supplementary Figure1). Yellow dotted line delineate somite boundary. The bar graph in Panel (E) shows the percentage of embryos with development defects. (F) RT-PCR analysis for confirmation of the efficacy of the gene X-MO. Left: RT-PCR of gene X transcript from uninjected and gene X-MO morpholino- injected embryos 2 days after fertilization, demonstrating skipping of exon 3. Right: Sanger sequencing of both the wild type band and the exon 3-skipped band validating the wild type sequence and the exon 3-skipped sequence.

Gene X fine-tunes Hedgehog signaling pathway (Quantitative Real-Time PCR)

Fig.2. Endogenous shha, ptch1, ptch2, sufu, gli1, gli2a, gli2b, and gli3 in wild-type control and gene X morphants assessed by qRT-PCR (n = 100 individual embryos). gene X fine-tunes Hedgehog patterning activity maybe through a novel regulatory feedback loop.

Morpholino knock down of gene X inhibits angiogenic vessel growth in zebrafish

Fig.3. gene X  knock down inhibits the trunk angiogenesis in zebrafish. (A-D) Representative fluorescent images of zebrafish embryos at 32h post-fertilization (hpf). (C-E) Compared with wild-type control, embryos injected with gene X-MO present a lower number of incomplete ISVs and only occasional sprouts (asterisk) of dorsal aorta. The boxed regions are shown at higher magnification in the right panels. DLAV, dorsal longitudinal anastomotic vessels; ISV, intersegmental vessels; DA, dorsal aorta; PCV, posterior cardinal vein.


Morpholino knock down of gene X induces CNS-specific apoptosis

Fig. 4. Morpholino knock down of gene X induces potent CNS-specific apoptosis. Wild-type control embryos and embryos injected with gene X-MO were stained with acridine orange (AO) at 32hpf. Apoptotic cells are visible as black spots, and less bright homogenous black staining is unspecific background staining. (A-B) Uninjected wild-type control zebrafish exhibited few or no apoptotic cells in CNS (central nervous system). In contrast, significantly increased staining was observed throughout  the CNS in zebrafish injected with gene X-MO (C-D). The blue boxed regions are shown at higher magnification in the right panels. A-D: lateral view, anterior, left.

Morpholino knock down of gene X induces potent ototoxicity in zebrafish

Fig.5. gene X knock down induces potent ototoxicity in zebrafish. Wild-type control embryos and embryos injected with gene X-MO were stained with the mitochondrial potentiometric dye DASPEI at 6-dpf. Hair cells stereotypically located on the lateral line were stained as green dots (white arrow). Uninjected wild-type control zebrafish exhibited normal hair cell number. In contrast, significantly decreased hair cell staining was observed in zebrafish injected with gene X-MO. Fluorescent DASPEI images were inverted for particle analysis. The fluorescence particle signal was quantified using morphometric analysis. dpf, days post fertilization.