FIGURE

Fig. 7

ID

ZDB-FIG-250423-39

Publication

Lin et al., 2025 - Optimizing gRNA selection for high-penetrance F0 CRISPR screening for interrogating disease gene function

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Fig. 7

Utilizing the F0 approach to investigate the tissue-specific effects of genes with transgenic reporter lines or through immunohistochemistry. A transgenic line, Tg(olig2:DsRed2;mnx1:EGFP), was used to reveal the morphology of (A) the cerebellum (dorsal view; n = 3 embryos) and (B) motor neurons (n = 5 embryos) with red fluorescent protein driven by the olig2 promoter. (C) Fluorescent conjugated phalloidin staining was used to reveal actin filaments in the skeletal muscles of the fish trunk at 10 dpf (n = 6 larvae). Blue arrowheads indicate missing (loosened) muscle fibers. (D) Alcian blue staining and Tg(col2a1a:EGFP-CAAX) was used to reveal the structures of cartilages in cog1 F0 knockouts at 5 dpf (n = 10 larvae). The magenta arrow indicates aberrant cartilage development. A ventral view is shown. (E) Bright-field images revealed the cardiac development of animals (n = 4 larvae). The heart was outlined by blue lines, and the blue arrow indicates empty space in pericardium. BuA, bulbus arteriosus; V, ventricle; and A, atrium are indicated. A ventral view is shown. (F) Blood vasculature was revealed by Tg(kdrl:EGFP) at 5 dpf (n = 6 larvae). DLAV, dorsal longitudinal anastomotic vessel; ISV, intersegmental vessel; and DA, dorsal aorta. White asterisks indicate aberrant connected vessel, white arrow indicates defective angiogenesis and the white arrowhead indicates enlarged ISV. (G) Oil Red O staining was used to visualize the lipid accumulation in animals at 8 dpf (n = 35 larvae). Animals are presented in lateral view, with anterior to the left, unless specifically addressed.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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