brunello knockout pooled lentiviral library  (Addgene inc)

Journal: Cell Reports Medicine

Article Title: Suppressing proteasome activity enhances sensitivity to actinomycin D in diffuse anaplastic Wilms tumor

doi: 10.1016/j.xcrm.2025.102133

Figure Lengend Snippet: ActD disrupts protein homeostasis in anaplastic Wilms tumor cells (A) Diagram of the effect of actD at the translational level. ActD depletes fully formed ribosomes and reduces translational capacity. The reduced ribosomes are distributed unevenly. Ribosome profiling provides a snapshot of transcripts actively undergoing translation. (B) Whole protein lysates of DMSO (vehicle)- or actD-treated WiT49 or 17.94, fluorescently labeled with AHA-AZDye 680 for nascent protein detection, and corresponding loading control (tubulin) detected by western blot from the same gel. (C and D) Top 5 most enriched and bottom 5 most depleted KEGG gene sets in ribosome profiling of actD- vs. DMSO-treated WiT49 after 6 h (C) or 72 h (D). (E) GSEA of genome-wide CRISPR knockout screen reveals top 10 most significant KEGG pathways that sensitize WiT49 to actD, ranked by p value. (F) Heatmap displaying RPPA results of actD- vs. DMSO-treated WiT49. Normalized, log2-transformed, median-centered values from validated antibodies with standard deviation over 0.1 are shown here.

Article Snippet: Human Brunello CRISPR knockout pooled library (Brunello) , Doench et al. , Addgene Pooled Library 73178.

Techniques: Wilms Tumor Assay, Labeling, Control, Western Blot, Genome Wide, CRISPR, Knock-Out, Transformation Assay, Standard Deviation