genome wide ko eko sgrna library  (Addgene inc)

Journal: Redox Biology

Article Title: CRISPR-based chemogenomic profiling reveals redox vulnerabilities to epigallocatechin-3-gallate and green tea polyphenol extract

doi: 10.1016/j.redox.2026.104047

Figure Lengend Snippet: Genome-wide CRISPR-Cas9 screen identifies genetic determinants of GTE-induced cytotoxicity. (A) Schematic overview of the experimental workflow for the genome-wide CRISPR knockout screen in NALM-6 cells treated with green tea extract (GTE), aimed at identifying genes modulating compound sensitivity. (B) Distribution of CRANKS scores for 19,034 genes following GTE treatment. Negative scores (red arrows, CRANKS ≤ −2) indicate sensitizer genes whose knockout enhances compound sensitivity, while positive scores (green arrows, CRANKS ≥2) represent rescuers whose knockout confers resistance. The 2D scatter plot is aligned with a one-dimensional frequency distribution, illustrating the thresholds used to define top hits. (C) Volcano plot showing CRANKS scores plotted against –log 10 p -values. Genes above the horizontal dashed line marks the p -threshold ( p < 0.05), while those beyond the vertical dashed lines exceed CRANKS thresholds of ±2, identifying 30 filtered genes ( p ≤ 0.01) labeled by gene symbol. Red points represent sensitizers; green points represent rescuers. Listed starred gene (∗) indicate top hits. (D) KEGG pathway enrichment analysis of highest CRANKS-filtered candidates (|CRANKS| ≥ 2; p ≤ 0.01; set including all top hits). Bubble color reflects false discovery rate (FDR), ranging from light green (FDR = 9 × 10 −7 ) to dark blue (FDR = 4 × 10 −3 ), and bubble size corresponds to the number of genes in each pathway. Pathways are ranked by significance on the y-axis. Analysis was performed using the STRING database with whole-genome background, requiring a minimum of two genes per pathway and FDR ≤0.05.

Article Snippet: Briefly, a human NALM-6 (pre-B ALL lymphocytes) clone bearing an integrated inducible Cas9 expression cassette generated by lentiviruses made from pCW-Cas9 (Addgene #50661) was transduced with the genome-wide KO EKO sgRNA library (278,754 different sgRNAs).

Techniques: Genome Wide, CRISPR, Knock-Out, Labeling

Journal: Redox Biology

Article Title: CRISPR-based chemogenomic profiling reveals redox vulnerabilities to epigallocatechin-3-gallate and green tea polyphenol extract

doi: 10.1016/j.redox.2026.104047

Figure Lengend Snippet: EGCG-induced pro-oxidant mechanisms: Insights from genome-wide CRISPR screening. (1) EGCG undergoes auto-oxidation generating reactive oxygen species (ROS) such as hydrogen peroxide (H 2 O 2 ) and quinone intermediates. A genome-wide CRISPR-Cas9 screen identified key genes modulating cellular response to EGCG-induced oxidative stress. (2) Knockouts of glutathione biosynthesis genes ( GCLC , GCLM , GSS ) impair H 2 O 2 detoxification, sensitizing cells to ferroptosis. (3) Peroxisomal genes ( PEX1 , PEX6 , PEX12 , PEX14 ) regulate ROS metabolism; their disruption compromises H 2 O 2 clearance via catalase ( CAT ) and peroxiredoxin-1 ( PRDX1 ). (4) The KEAP1-NRF2 axis controls antioxidant gene expression, inducing enzymes (e.g., CAT, PRDX1, GCLs) that mitigate ROS toxicity. KEAP1 knockout enhances NRF2 signaling and confers resistance. (5) Additional modulators include ABCC1 (drug efflux), SLC7A11 (cysteine transporter) and BAK1 (apoptosis). Sensitizer hits (red labels) indicate knockouts that heighten EGCG toxicity, while resistance hits (green labels) protect against cell death. Color intensity reflects CRANKS scores relative to the highest scoring genes. Together, these findings illustrate how EGCG's pro-oxidant activity can overwhelm cancer cell defenses when redox-regulating pathways are genetically compromised.

Article Snippet: Briefly, a human NALM-6 (pre-B ALL lymphocytes) clone bearing an integrated inducible Cas9 expression cassette generated by lentiviruses made from pCW-Cas9 (Addgene #50661) was transduced with the genome-wide KO EKO sgRNA library (278,754 different sgRNAs).

Techniques: Genome Wide, CRISPR, Disruption, Gene Expression, Knock-Out, Activity Assay