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Addgene inc human rbp pooled crispr ko library
Human Rbp Pooled Crispr Ko Library, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+crispr+ko+pooled+library/pm41854249-84-10-20?v=Addgene+inc
Average 93 stars, based on 5 article reviews

human rbp pooled crispr ko library - by Bioz Stars, 2026-06

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a , Schematic of CRISPR–Cas9 knock-in strategy. A C-terminal GFP-P2A-BFP tag was inserted into the FSP1 genomic locus before the stop codon. Cells expressing both GFP and BFP were enriched through multiple rounds of FACS. b , Fluorescence histograms of WT control and genomic FSP1 GFP-P2A-BFP knock-in cells. c , Immunoblot of lysates from control and FSP1 GFP-P2A-BFP cells from b . d , Fluorescence histograms of FSP1 GFP-P2A-BFP cells incubated with siRNAs against FSP1 for 72 h, including siRNAs from scramble control. e , Immunoblot of control and FSP1 GFP-P2A-BFP cells from d . f , FSP1 GFP-P2A-BFP subcellular distribution by live-cell microscopy. Cells were treated with 200 µM oleate for 24 h to induce lipid droplet formation, 500 nM LipiBlue to label lipid droplets and 5 µg ml −1 CellMask deep red to label the plasma membrane. g , Dose response of RSL3-induced cell death of WT and FSP1 GFP-P2A-BFP reporter cells in the presence or absence of 2 µM FSEN1 for 24 h. Data are shown as the mean ± s.e.m. of three wells of a 96-well plate from three independent experiments. h , Fluorescence histograms of FSP1 GFP-P2A-BFP cells treated with proteolysis inhibitors (5 mM 3-MA, 250 nM Baf-A1, 5 µM CB5083, 10 µM MLN7243 and 10 µM MG132) for 6 h. i , Quantification of MFI change in GFP:BFP ratio from h . Data are shown as the median ± s.e.m. of four independent experiments. NS, not significant.

Journal: Nature Structural & Molecular Biology

Article Title: Vitamin B2 metabolism promotes FSP1 stability to prevent ferroptosis

doi: 10.1038/s41594-026-01759-x

Figure Lengend Snippet: a , Schematic of CRISPR–Cas9 knock-in strategy. A C-terminal GFP-P2A-BFP tag was inserted into the FSP1 genomic locus before the stop codon. Cells expressing both GFP and BFP were enriched through multiple rounds of FACS. b , Fluorescence histograms of WT control and genomic FSP1 GFP-P2A-BFP knock-in cells. c , Immunoblot of lysates from control and FSP1 GFP-P2A-BFP cells from b . d , Fluorescence histograms of FSP1 GFP-P2A-BFP cells incubated with siRNAs against FSP1 for 72 h, including siRNAs from scramble control. e , Immunoblot of control and FSP1 GFP-P2A-BFP cells from d . f , FSP1 GFP-P2A-BFP subcellular distribution by live-cell microscopy. Cells were treated with 200 µM oleate for 24 h to induce lipid droplet formation, 500 nM LipiBlue to label lipid droplets and 5 µg ml −1 CellMask deep red to label the plasma membrane. g , Dose response of RSL3-induced cell death of WT and FSP1 GFP-P2A-BFP reporter cells in the presence or absence of 2 µM FSEN1 for 24 h. Data are shown as the mean ± s.e.m. of three wells of a 96-well plate from three independent experiments. h , Fluorescence histograms of FSP1 GFP-P2A-BFP cells treated with proteolysis inhibitors (5 mM 3-MA, 250 nM Baf-A1, 5 µM CB5083, 10 µM MLN7243 and 10 µM MG132) for 6 h. i , Quantification of MFI change in GFP:BFP ratio from h . Data are shown as the median ± s.e.m. of four independent experiments. NS, not significant.

Article Snippet: Genome-wide CRISPR–Cas9 screens were performed using the Bassik human CRISPR KO library (Addgene, pooled libraries 101926–101934).

Techniques: CRISPR, Knock-In, Expressing, Fluorescence, Control, Western Blot, Incubation, Microscopy, Clinical Proteomics, Membrane

a , Schematic of the genome-wide CRISPR–Cas9 screening strategy. b , Gene effects and gene scores calculated for individual genes analyzed in the genome-wide CRISPR screen. c , Top, cloud plot indicating count numbers corresponding to FSP1 (color scale) and control (gray scale) sgRNAs. Bottom, distribution for the relative enrichment of FSP1 sgRNAs (blue lines). The gray line indicates the mean of the control sgRNA distribution. d , Bubble plot of the genes identified from the genome-wide screen with a 10% FDR cutoff, where each bubble color represents an enriched GO term functional cluster. e , Scatter plot of signed gene scores for individual genes from genome-wide screen ( x axis) versus batch retest screen ( y axis).

Journal: Nature Structural & Molecular Biology

Article Title: Vitamin B2 metabolism promotes FSP1 stability to prevent ferroptosis

doi: 10.1038/s41594-026-01759-x

Figure Lengend Snippet: a , Schematic of the genome-wide CRISPR–Cas9 screening strategy. b , Gene effects and gene scores calculated for individual genes analyzed in the genome-wide CRISPR screen. c , Top, cloud plot indicating count numbers corresponding to FSP1 (color scale) and control (gray scale) sgRNAs. Bottom, distribution for the relative enrichment of FSP1 sgRNAs (blue lines). The gray line indicates the mean of the control sgRNA distribution. d , Bubble plot of the genes identified from the genome-wide screen with a 10% FDR cutoff, where each bubble color represents an enriched GO term functional cluster. e , Scatter plot of signed gene scores for individual genes from genome-wide screen ( x axis) versus batch retest screen ( y axis).

Article Snippet: Genome-wide CRISPR–Cas9 screens were performed using the Bassik human CRISPR KO library (Addgene, pooled libraries 101926–101934).

Techniques: Genome Wide, CRISPR, Control, Functional Assay

a , Schematic of the CRISPR–Cas9 batch retest screening strategy. b , c , Gene effects and gene scores calculated for individual genes analyzed in the batch retest CRISPR screens for expression (BFP; b ) or stability (GFP:BFP; c ). d , Heat maps of clustered genes based on the signed gene scores from GO enrichment analysis of the core transcriptional regulators. e , Schematic of cytosolic redox pathways including KEAP1 and NFE2L2 signaling, selenocysteine synthesis, CoQ synthesis and glycolysis. Genes are annotated with modes corresponding to gene effects and scores from batch retest of untreated and 100 nM RSL3-treated expression (BFP) screens. Red asterisks indicate genes reported to be regulated by NFE2L2. f , Heat maps of clustered genes based on the signed gene scores from GO enrichment analysis of the core post-translational regulators.

Journal: Nature Structural & Molecular Biology

Article Title: Vitamin B2 metabolism promotes FSP1 stability to prevent ferroptosis

doi: 10.1038/s41594-026-01759-x

Figure Lengend Snippet: a , Schematic of the CRISPR–Cas9 batch retest screening strategy. b , c , Gene effects and gene scores calculated for individual genes analyzed in the batch retest CRISPR screens for expression (BFP; b ) or stability (GFP:BFP; c ). d , Heat maps of clustered genes based on the signed gene scores from GO enrichment analysis of the core transcriptional regulators. e , Schematic of cytosolic redox pathways including KEAP1 and NFE2L2 signaling, selenocysteine synthesis, CoQ synthesis and glycolysis. Genes are annotated with modes corresponding to gene effects and scores from batch retest of untreated and 100 nM RSL3-treated expression (BFP) screens. Red asterisks indicate genes reported to be regulated by NFE2L2. f , Heat maps of clustered genes based on the signed gene scores from GO enrichment analysis of the core post-translational regulators.

Article Snippet: Genome-wide CRISPR–Cas9 screens were performed using the Bassik human CRISPR KO library (Addgene, pooled libraries 101926–101934).

Techniques: CRISPR, Expressing

a , Schematic of sensitized UBAL degradation CRISPR–Cas9 screen. b , Gene effects and gene scores calculated for individual genes analyzed in the targeted degradation screen. c , STRING network map of the identified genes with a 10% FDR cutoff, where each bubble color represents an enriched GO term functional cluster. d , Fluorescence histograms of FSP1 KO cells expressing FSP1–GFP in the indicated cell lines. e , Quantification of MFI change in GFP from d . Data are shown as the median ± s.e.m. of four independent experiments. f , Immunoblot of lysates from d . g , h , Ubiquitinated FSP1–GFP conjugates affinity-captured from lysates treated with 1 µM MG132 for 24 h ( g ) or expressing exogenous RNF8 variants ( h ). i , Kinetics of GFP fluorescence decay of FSP1 KO cells expressing FSP1–GFP in Cas9 control or RFK KO cells with or without the loss of RNF8 following Dox washout. j , FSP1–GFP or RNF8 S-tag immunoprecipitation from FSP1 KO cells expressing FSP1–GFP in the indicated cell lines and blotting for RNF8 (top) or FSP1 (middle). k , Model illustrating the mechanism by which vitamin B2 metabolism and FAD synthesis impact FSP1 activity and stability to prevent ferroptosis.

Journal: Nature Structural & Molecular Biology

Article Title: Vitamin B2 metabolism promotes FSP1 stability to prevent ferroptosis

doi: 10.1038/s41594-026-01759-x

Figure Lengend Snippet: a , Schematic of sensitized UBAL degradation CRISPR–Cas9 screen. b , Gene effects and gene scores calculated for individual genes analyzed in the targeted degradation screen. c , STRING network map of the identified genes with a 10% FDR cutoff, where each bubble color represents an enriched GO term functional cluster. d , Fluorescence histograms of FSP1 KO cells expressing FSP1–GFP in the indicated cell lines. e , Quantification of MFI change in GFP from d . Data are shown as the median ± s.e.m. of four independent experiments. f , Immunoblot of lysates from d . g , h , Ubiquitinated FSP1–GFP conjugates affinity-captured from lysates treated with 1 µM MG132 for 24 h ( g ) or expressing exogenous RNF8 variants ( h ). i , Kinetics of GFP fluorescence decay of FSP1 KO cells expressing FSP1–GFP in Cas9 control or RFK KO cells with or without the loss of RNF8 following Dox washout. j , FSP1–GFP or RNF8 S-tag immunoprecipitation from FSP1 KO cells expressing FSP1–GFP in the indicated cell lines and blotting for RNF8 (top) or FSP1 (middle). k , Model illustrating the mechanism by which vitamin B2 metabolism and FAD synthesis impact FSP1 activity and stability to prevent ferroptosis.

Article Snippet: Genome-wide CRISPR–Cas9 screens were performed using the Bassik human CRISPR KO library (Addgene, pooled libraries 101926–101934).

Techniques: CRISPR, Functional Assay, Fluorescence, Expressing, Western Blot, Control, Immunoprecipitation, Activity Assay

ACE2-independent binding of the Omicron RBD to cell surface HS. ( A ) CRISPR KO library screening scheme to identify Omicron RBD binding molecules expressed on HEK293T cells. ( B ) Binding of CRISPR KO library-transduced HEK293T cells with (red line) or without (shaded gray) Omicron RBD-Fc, before and after sorting. ( C ) Number of sgRNA sequences identified in Omicron RBD-Fc non-binding cells after sorting. Red: GAG-synthesis-related genes; black: other expressed genes; gray: non-expressed genes in HEK293T cells. ( D ) HS biosynthetic pathway highlighting SLC35B2 and B3GAT3. PAPS, 3'-phosphoadenosine-5'-phosphosulfate; Xyl, xylose; Gal, galactose; GlcNAc, N-acetylglucosamine; GlcA, glucuronic acid; IdoA, iduronic acid. ( E ) Binding of BA.1 RBD-Fc, anti-HS Ab, or anti-CS Ab to mock (black line), or SLC35B2 or B3GAT3 KO (red line) HEK293T cells. ( F ) Binding of WT or BA.1 RBD-Fc, PILRα-Fc, anti-HS Ab, or anti-CS Ab to ACE2 KO HEK293T cells pretreated with (+) or without (–) heparinase or chondroitinase. ( G ) Binding of WT or BA.1 RBD-Fc to HEK293T cells at different heparin concentrations. RBD-Fc binding was normalized to binding in the absence of heparin. ( H ) Immunofluorescence of human nasal tissue with anti-HS Ab and 4', 6-diamidino-2-phenylindole (DAPI). Scale bar, 200 µm. ( I ) Binding of WT or BA.1 RBD-Fc, anti-ACE2 Ab, or anti-HS Ab to cell lines. Data are mean ± SEM of three to four technical replicates. Statistical analysis was performed using two-way ANOVA with Sidak’s multiple comparison tests in panel F ; * P < 0.05, ** P < 0.01, and **** P < 0.0001; ns, not significant. Data are representative of two to three independent experiments.

Journal: mBio

Article Title: Evolutionary dynamics of heparan sulfate utilization by SARS-CoV-2

doi: 10.1128/mbio.01303-25

Figure Lengend Snippet: ACE2-independent binding of the Omicron RBD to cell surface HS. ( A ) CRISPR KO library screening scheme to identify Omicron RBD binding molecules expressed on HEK293T cells. ( B ) Binding of CRISPR KO library-transduced HEK293T cells with (red line) or without (shaded gray) Omicron RBD-Fc, before and after sorting. ( C ) Number of sgRNA sequences identified in Omicron RBD-Fc non-binding cells after sorting. Red: GAG-synthesis-related genes; black: other expressed genes; gray: non-expressed genes in HEK293T cells. ( D ) HS biosynthetic pathway highlighting SLC35B2 and B3GAT3. PAPS, 3'-phosphoadenosine-5'-phosphosulfate; Xyl, xylose; Gal, galactose; GlcNAc, N-acetylglucosamine; GlcA, glucuronic acid; IdoA, iduronic acid. ( E ) Binding of BA.1 RBD-Fc, anti-HS Ab, or anti-CS Ab to mock (black line), or SLC35B2 or B3GAT3 KO (red line) HEK293T cells. ( F ) Binding of WT or BA.1 RBD-Fc, PILRα-Fc, anti-HS Ab, or anti-CS Ab to ACE2 KO HEK293T cells pretreated with (+) or without (–) heparinase or chondroitinase. ( G ) Binding of WT or BA.1 RBD-Fc to HEK293T cells at different heparin concentrations. RBD-Fc binding was normalized to binding in the absence of heparin. ( H ) Immunofluorescence of human nasal tissue with anti-HS Ab and 4', 6-diamidino-2-phenylindole (DAPI). Scale bar, 200 µm. ( I ) Binding of WT or BA.1 RBD-Fc, anti-ACE2 Ab, or anti-HS Ab to cell lines. Data are mean ± SEM of three to four technical replicates. Statistical analysis was performed using two-way ANOVA with Sidak’s multiple comparison tests in panel F ; * P < 0.05, ** P < 0.01, and **** P < 0.0001; ns, not significant. Data are representative of two to three independent experiments.

Article Snippet: CRISPR KO library-transduced HEK293T cells were generated by lentivirus-mediated transduction using the Human CRISPR KO Pooled Library (GeCKO v2; Addgene, 1000000048).

Techniques: Binding Assay, CRISPR, Library Screening, Immunofluorescence, Comparison

A kinase domain–targeted CRISPR screen identifies HASPIN as a novel dependency in AML cells. (A) Schematic of human kinase domain–targeted CRISPR screen in 2 t(8;21) AML cell lines, Kasumi-1 and SKNO-1. (B) Gene rank plots (left) and volcano plots (right) depicting significant kinase hits in the kinase domain–targeted CRISPR screens. Top candidates determined by CRISPR score as calculated by MAGeCK robust ranking aggregation (RRA) (left) and a log 2 (fold change) ≤−1.0 and false discovery rate (FDR) ≤0.05 significance cutoff (right). White diamonds indicate top 10 kinase hits in each plot. (C) Bubble plot of preranked gene set enrichment analysis results performed on top kinases identified by CRISPR screen in Kasumi-1 and SKNO-1 AML cell lines. Fill color indicates normalized enrichment score (NES). Size indicates significance by –log 10 (FDR). Facets indicate Molecular Signatures Database (MSigDB) gene set collection. (D) Density plot of all individual sgRNA log 2 (fold change) values in the kinase domain–targeted library. For selected genes, log 2 (fold change) values of corresponding sgRNAs depicted for Kasumi-1 and SKNO-1 cell lines relative to all other library sgRNAs (red, blue, and gray, respectively). (E) Competitive proliferation assay of Kasumi-1 or SKNO-1 cells expressing nontargeting negative control, RPA3-targeting positive control, or 1 of 2 HASPIN-targeting sgRNAs derived from CRISPR screen. Relative changes in cell proliferation rate measured by percentage of GFP-positive cells relative to nontargeting control on each day. Data are mean ± standard deviation (SD) of 4 independent experiments. CGP, chemical, genetic perturbation; CP, canonical pathway.

Journal: Blood Neoplasia

Article Title: Targeting HASPIN kinase disrupts SR protein–mediated RNA splicing and synergizes with BCL-2 inhibitor venetoclax in AML

doi: 10.1016/j.bneo.2025.100107

Figure Lengend Snippet: A kinase domain–targeted CRISPR screen identifies HASPIN as a novel dependency in AML cells. (A) Schematic of human kinase domain–targeted CRISPR screen in 2 t(8;21) AML cell lines, Kasumi-1 and SKNO-1. (B) Gene rank plots (left) and volcano plots (right) depicting significant kinase hits in the kinase domain–targeted CRISPR screens. Top candidates determined by CRISPR score as calculated by MAGeCK robust ranking aggregation (RRA) (left) and a log 2 (fold change) ≤−1.0 and false discovery rate (FDR) ≤0.05 significance cutoff (right). White diamonds indicate top 10 kinase hits in each plot. (C) Bubble plot of preranked gene set enrichment analysis results performed on top kinases identified by CRISPR screen in Kasumi-1 and SKNO-1 AML cell lines. Fill color indicates normalized enrichment score (NES). Size indicates significance by –log 10 (FDR). Facets indicate Molecular Signatures Database (MSigDB) gene set collection. (D) Density plot of all individual sgRNA log 2 (fold change) values in the kinase domain–targeted library. For selected genes, log 2 (fold change) values of corresponding sgRNAs depicted for Kasumi-1 and SKNO-1 cell lines relative to all other library sgRNAs (red, blue, and gray, respectively). (E) Competitive proliferation assay of Kasumi-1 or SKNO-1 cells expressing nontargeting negative control, RPA3-targeting positive control, or 1 of 2 HASPIN-targeting sgRNAs derived from CRISPR screen. Relative changes in cell proliferation rate measured by percentage of GFP-positive cells relative to nontargeting control on each day. Data are mean ± standard deviation (SD) of 4 independent experiments. CGP, chemical, genetic perturbation; CP, canonical pathway.

Article Snippet: CRISPR screen was performed using the human kinase domain–focused CRISPR knockout (KO) library (Addgene 117725; a gift from Christopher Vakoc) and 2 t(8;21) AML cell lines.

Techniques: CRISPR, Proliferation Assay, Expressing, Negative Control, Positive Control, Derivative Assay, Control, Standard Deviation

HASPIN is a clinically relevant, general leukemia dependency. (A) Bar plot depicting mean log 2 (fold change) of HASPIN targeting sgRNA genome-wide CRISPR screen performed in several leukemia cell lines as reported by Wang et al. Screen data were retrieved from BIOGRID ORCS. Dotted line indicates author-specified significance cutoff. (B) Competitive proliferation assay of THP-1 or OCI-AML3 cells expressing nontargeting negative control, RPA3-targeting positive control, or 1 of 2 HASPIN-targeting sgRNAs derived from CRISPR screen. Relative changes in cell proliferation rate measured by percentage of GFP-positive cells relative to nontargeting control on each day. Data are mean ± SD of 4 independent experiments per cell line. (C) Box plots depicting median HASPIN mRNA expression in the TCGA-LAML patient cohort separated by AML subtype. MLL (KMT2A) or RUNX1-RUNX1T1 t(8;21) translocation cohorts are highlighted in green and orange, respectively. Individuals with KMT2A structural variants are indicated with purple diamonds. (D) Box plots depicting median HASPIN mRNA expression in the BEAT-AML (2022) patient cohort separated by AML subtype. MLL (KMT2A) or RUNX1-RUNX1T1 t(8;21) translocation cohorts are highlighted in green and orange, respectively. Individuals with KMT2A structural variants are indicated with purple diamonds. (E) Kaplan-Meier survival curve depicting comparison of overall survival of patients with TCGA-LAML belonging to the top quartile (red) and bottom quartile (blue) of HASPIN expression. Plot and data derived from GEPIA2. (F) Forest plot of hazard ratios from multivariate Cox proportional hazard analysis of overall survival of patients with TCGA LAML incorporating HASPIN expression level and significant clinical and genetic factors. High and low HASPIN -expressing patients belong to the top and bottom expression quartiles, respectively. Clinical variables include the following: patient sex (Sex), age at first diagnosis (Diagnosis_Age), genetic risk group (Risk_Group), FLT3 mutation status (FLT3_Status), NPM1 mutation status (NPM1_Status), DNMT3A mutation status (DNMT3A_Status), TP53 mutation status (TP53_Status), and NRAS mutation status (NRAS_Status). Clinical metadata and mutation calls derived from the Genomic Data Commons TCGA LAML project patient information. N.D., not defined; NOS, not otherwise specified; NP, not profiled.

Journal: Blood Neoplasia

Article Title: Targeting HASPIN kinase disrupts SR protein–mediated RNA splicing and synergizes with BCL-2 inhibitor venetoclax in AML

doi: 10.1016/j.bneo.2025.100107

Figure Lengend Snippet: HASPIN is a clinically relevant, general leukemia dependency. (A) Bar plot depicting mean log 2 (fold change) of HASPIN targeting sgRNA genome-wide CRISPR screen performed in several leukemia cell lines as reported by Wang et al. Screen data were retrieved from BIOGRID ORCS. Dotted line indicates author-specified significance cutoff. (B) Competitive proliferation assay of THP-1 or OCI-AML3 cells expressing nontargeting negative control, RPA3-targeting positive control, or 1 of 2 HASPIN-targeting sgRNAs derived from CRISPR screen. Relative changes in cell proliferation rate measured by percentage of GFP-positive cells relative to nontargeting control on each day. Data are mean ± SD of 4 independent experiments per cell line. (C) Box plots depicting median HASPIN mRNA expression in the TCGA-LAML patient cohort separated by AML subtype. MLL (KMT2A) or RUNX1-RUNX1T1 t(8;21) translocation cohorts are highlighted in green and orange, respectively. Individuals with KMT2A structural variants are indicated with purple diamonds. (D) Box plots depicting median HASPIN mRNA expression in the BEAT-AML (2022) patient cohort separated by AML subtype. MLL (KMT2A) or RUNX1-RUNX1T1 t(8;21) translocation cohorts are highlighted in green and orange, respectively. Individuals with KMT2A structural variants are indicated with purple diamonds. (E) Kaplan-Meier survival curve depicting comparison of overall survival of patients with TCGA-LAML belonging to the top quartile (red) and bottom quartile (blue) of HASPIN expression. Plot and data derived from GEPIA2. (F) Forest plot of hazard ratios from multivariate Cox proportional hazard analysis of overall survival of patients with TCGA LAML incorporating HASPIN expression level and significant clinical and genetic factors. High and low HASPIN -expressing patients belong to the top and bottom expression quartiles, respectively. Clinical variables include the following: patient sex (Sex), age at first diagnosis (Diagnosis_Age), genetic risk group (Risk_Group), FLT3 mutation status (FLT3_Status), NPM1 mutation status (NPM1_Status), DNMT3A mutation status (DNMT3A_Status), TP53 mutation status (TP53_Status), and NRAS mutation status (NRAS_Status). Clinical metadata and mutation calls derived from the Genomic Data Commons TCGA LAML project patient information. N.D., not defined; NOS, not otherwise specified; NP, not profiled.

Article Snippet: CRISPR screen was performed using the human kinase domain–focused CRISPR knockout (KO) library (Addgene 117725; a gift from Christopher Vakoc) and 2 t(8;21) AML cell lines.

Techniques: Genome Wide, CRISPR, Proliferation Assay, Expressing, Negative Control, Positive Control, Derivative Assay, Control, Translocation Assay, Comparison, Biomarker Discovery, Mutagenesis

HASPIN inhibitor CHR-6494 effectively targets AML and synergizes with BCL-2 inhibition. (A) Dose-response curves (left) and IC comparison (right) of Kasumi-1 and healthy CD34 + hematopoietic progenitor cells treated with CHR-6494. IC values determined by nonlinear regression. Data on curve are mean ± SD of technical triplicates. Representative curves of 3 independent experiments revealed. Data on bar plot are mean ± SD of 3 independent experiments. Significance determined by unpaired 2-tailed Student t test. ∗∗∗∗ P < .0001. (B) Bar plots comparing CHR-6494 IC values in leukemia cell lines. IC values determined by dose-response curve with nonlinear regression for each cell line. Data are mean ± SD of 3 independent experiments. Dotted line indicates CHR-6494 IC value of healthy CD34 + hematopoietic progenitor cells determined in panel A. (C) Bar plots depicting normalized HASPIN sgRNA counts in a genome-wide CRISPR screen in MOLM-13 cells treated with either DMSO or VEN for 8 or 16 days as performed by Chen et al. Screen data were retrieved from BIOGRID ORCS. Counts were normalized to initial time point (d0). One data point was removed from DMSO (d16) as a significant outlier. Data are mean ± SD. Significance determined by 1-way ANOVA with Holm-Sidak multiple comparison correction. ∗ P < .05; ∗∗ P < .01. (D) Dose-response matrix (left) and corresponding zero interaction potency (ZIP) drug synergy contour plot (right) of Kasumi-1 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. (E) Dose-response matrix (left) and corresponding ZIP drug synergy contour plot (right) of THP-1 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. (F) Dose-response matrix (left) and corresponding ZIP drug synergy contour plot (right) of OCI-AML3 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. DMSO, dimethyl sulfoxide.

Journal: Blood Neoplasia

Article Title: Targeting HASPIN kinase disrupts SR protein–mediated RNA splicing and synergizes with BCL-2 inhibitor venetoclax in AML

doi: 10.1016/j.bneo.2025.100107

Figure Lengend Snippet: HASPIN inhibitor CHR-6494 effectively targets AML and synergizes with BCL-2 inhibition. (A) Dose-response curves (left) and IC comparison (right) of Kasumi-1 and healthy CD34 + hematopoietic progenitor cells treated with CHR-6494. IC values determined by nonlinear regression. Data on curve are mean ± SD of technical triplicates. Representative curves of 3 independent experiments revealed. Data on bar plot are mean ± SD of 3 independent experiments. Significance determined by unpaired 2-tailed Student t test. ∗∗∗∗ P < .0001. (B) Bar plots comparing CHR-6494 IC values in leukemia cell lines. IC values determined by dose-response curve with nonlinear regression for each cell line. Data are mean ± SD of 3 independent experiments. Dotted line indicates CHR-6494 IC value of healthy CD34 + hematopoietic progenitor cells determined in panel A. (C) Bar plots depicting normalized HASPIN sgRNA counts in a genome-wide CRISPR screen in MOLM-13 cells treated with either DMSO or VEN for 8 or 16 days as performed by Chen et al. Screen data were retrieved from BIOGRID ORCS. Counts were normalized to initial time point (d0). One data point was removed from DMSO (d16) as a significant outlier. Data are mean ± SD. Significance determined by 1-way ANOVA with Holm-Sidak multiple comparison correction. ∗ P < .05; ∗∗ P < .01. (D) Dose-response matrix (left) and corresponding zero interaction potency (ZIP) drug synergy contour plot (right) of Kasumi-1 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. (E) Dose-response matrix (left) and corresponding ZIP drug synergy contour plot (right) of THP-1 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. (F) Dose-response matrix (left) and corresponding ZIP drug synergy contour plot (right) of OCI-AML3 cells treated with CHR-6494 and VEN combination for 48 hours. Each cell represents drug combined at indicated concentrations. Treatment response is percent inhibition; higher values indicate lower cell viability. Synergy scores represent ZIP synergy calculations of inhibition effects exceeding values expected between 2 noninteracting agents. Mean synergy scores and significance reported at top of respective contour plot. Representative plots of 3 independent experiments revealed. DMSO, dimethyl sulfoxide.

Article Snippet: CRISPR screen was performed using the human kinase domain–focused CRISPR knockout (KO) library (Addgene 117725; a gift from Christopher Vakoc) and 2 t(8;21) AML cell lines.

Techniques: Inhibition, Comparison, Genome Wide, CRISPR

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