Journal: bioRxiv
Article Title: YAP disrupts bile acid homeostasis to drive cancer-associated cachexia
doi: 10.64898/2026.02.01.702698
Figure Lengend Snippet: a , Schematic of the CRISPR–Cas9 knockout strategy targeting slc45a2 , designed to induce a visible pigmentation-loss phenotype for screening. Representative brightfield images of 6 and 21 dpf WT and cr slc45a2 zebrafish show loss of melanin pigmentation in mutants. b , Schematic of CRISPR guide RNA target sites (aa and ab) on the slc45a2 gene with representative Sanger sequencing traces and ICE analysis of indels from 7 dpf larvae injected at the one-cell stage with both guides. c , Indel efficiency rates for each slc45a2 CRISPR target site. d , Schematic of CRISPR guide RNA target sites (aa and ab) on the tgr5 gene with representative Sanger sequencing traces and ICE analysis from 7 dpf larvae co-injected at the one-cell stage with tgr5 and slc45a2 guides (latter not shown). e , Indel efficiency rates for each tgr5 CRISPR target site. f , Schematic of the CRISPR target site (ab) on cyp7a1 with corresponding Sanger sequencing traces and ICE analysis from 7 dpf larvae co-injected at the one-cell stage with cyp7a1 and slc45a2 guides (latter not shown). g , Indel efficiency rates for the cyp7a1 CRISPR target site. h-k , Biometrics of body length ( h , standard length, SL), muscle width ( i ), liver size ( j ) and adipose tissue size ( k ) in WT cr slc45a2 , WT cr tgr5 and WT cr cyp7a1 zebrafish at 21 dpf. l , Heat map of selected adipogenic, lipolytic, myogenic, proteolytic and inflammatory gene expression profiles in pooled WT, crslc45a2 , crtgr5 and crcyp7a1 zebrafish at 21 dpf, determined by bulk RNA-seq (n = 4 samples of 5 fish per condition). All data are mean ± s.e.m. P values were determined by one-way ANOVA with Tukey’s multiple-comparisons test ( h,i ) and Kruskal–Wallis test with Dunn’s multiple-comparisons post hoc adjustment ( j,k ). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.
Article Snippet: Indels in the sequenced data were analysed using the Inference of CRISPR Edits (ICE) web-based platform (Synthego).
Techniques: CRISPR, Knock-Out, Sequencing, Injection, Gene Expression, RNA Sequencing
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