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<t>CRISPR</t> screening revealed that co‐targeting WEE1 and AKT as a potential combination strategy in ARID1A/TP53 concurrent mutant CRC. A) Schematic description of the CRISPR knockout <t>kinome</t> library screen. (B) Results of CRISPR screen in SNUC5 cells. Scatter plots present the Z‐scores of average log 2 (fold change) for MK‐1775 vs DMSO ( x ‐axis) and for ZN‐c3 vs DMSO ( y ‐axis). (C) Immunoblot analysis confirming AKT2 knockout in SNUC5 cells (n=2). (D) Dose‐response curves of SNUC5 cells, treated with MK‐1775 (left) or ZN‐c3 (right) for 5 days. The results are representative of three independent experiments, each done in triplicate. Bars represent mean ± SD. (E) Immunoblot analysis of HCT15, SNUC5, COLO205, and HRT18 cell lines treated with 0.25 or 0.5 µ m MK‐1775 (n = 2). (F) GDSC2 analysis of indicated PI3K or AKT inhibitors according to ARID1A mutation status. Data are presented as scatter plots. Mean values are indicated by red lines. * p < 0.05, ** p < 0.01 (two‐tailed Wilcoxon rank‐sum test).
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<t>CRISPR</t> screening revealed that co‐targeting WEE1 and AKT as a potential combination strategy in ARID1A/TP53 concurrent mutant CRC. A) Schematic description of the CRISPR knockout <t>kinome</t> library screen. (B) Results of CRISPR screen in SNUC5 cells. Scatter plots present the Z‐scores of average log 2 (fold change) for MK‐1775 vs DMSO ( x ‐axis) and for ZN‐c3 vs DMSO ( y ‐axis). (C) Immunoblot analysis confirming AKT2 knockout in SNUC5 cells (n=2). (D) Dose‐response curves of SNUC5 cells, treated with MK‐1775 (left) or ZN‐c3 (right) for 5 days. The results are representative of three independent experiments, each done in triplicate. Bars represent mean ± SD. (E) Immunoblot analysis of HCT15, SNUC5, COLO205, and HRT18 cell lines treated with 0.25 or 0.5 µ m MK‐1775 (n = 2). (F) GDSC2 analysis of indicated PI3K or AKT inhibitors according to ARID1A mutation status. Data are presented as scatter plots. Mean values are indicated by red lines. * p < 0.05, ** p < 0.01 (two‐tailed Wilcoxon rank‐sum test).
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<t>CRISPR</t> screening revealed that co‐targeting WEE1 and AKT as a potential combination strategy in ARID1A/TP53 concurrent mutant CRC. A) Schematic description of the CRISPR knockout <t>kinome</t> library screen. (B) Results of CRISPR screen in SNUC5 cells. Scatter plots present the Z‐scores of average log 2 (fold change) for MK‐1775 vs DMSO ( x ‐axis) and for ZN‐c3 vs DMSO ( y ‐axis). (C) Immunoblot analysis confirming AKT2 knockout in SNUC5 cells (n=2). (D) Dose‐response curves of SNUC5 cells, treated with MK‐1775 (left) or ZN‐c3 (right) for 5 days. The results are representative of three independent experiments, each done in triplicate. Bars represent mean ± SD. (E) Immunoblot analysis of HCT15, SNUC5, COLO205, and HRT18 cell lines treated with 0.25 or 0.5 µ m MK‐1775 (n = 2). (F) GDSC2 analysis of indicated PI3K or AKT inhibitors according to ARID1A mutation status. Data are presented as scatter plots. Mean values are indicated by red lines. * p < 0.05, ** p < 0.01 (two‐tailed Wilcoxon rank‐sum test).
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<t>CRISPR</t> screening revealed that co‐targeting WEE1 and AKT as a potential combination strategy in ARID1A/TP53 concurrent mutant CRC. A) Schematic description of the CRISPR knockout <t>kinome</t> library screen. (B) Results of CRISPR screen in SNUC5 cells. Scatter plots present the Z‐scores of average log 2 (fold change) for MK‐1775 vs DMSO ( x ‐axis) and for ZN‐c3 vs DMSO ( y ‐axis). (C) Immunoblot analysis confirming AKT2 knockout in SNUC5 cells (n=2). (D) Dose‐response curves of SNUC5 cells, treated with MK‐1775 (left) or ZN‐c3 (right) for 5 days. The results are representative of three independent experiments, each done in triplicate. Bars represent mean ± SD. (E) Immunoblot analysis of HCT15, SNUC5, COLO205, and HRT18 cell lines treated with 0.25 or 0.5 µ m MK‐1775 (n = 2). (F) GDSC2 analysis of indicated PI3K or AKT inhibitors according to ARID1A mutation status. Data are presented as scatter plots. Mean values are indicated by red lines. * p < 0.05, ** p < 0.01 (two‐tailed Wilcoxon rank‐sum test).
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CRISPR screening revealed that co‐targeting WEE1 and AKT as a potential combination strategy in ARID1A/TP53 concurrent mutant CRC. A) Schematic description of the CRISPR knockout kinome library screen. (B) Results of CRISPR screen in SNUC5 cells. Scatter plots present the Z‐scores of average log 2 (fold change) for MK‐1775 vs DMSO ( x ‐axis) and for ZN‐c3 vs DMSO ( y ‐axis). (C) Immunoblot analysis confirming AKT2 knockout in SNUC5 cells (n=2). (D) Dose‐response curves of SNUC5 cells, treated with MK‐1775 (left) or ZN‐c3 (right) for 5 days. The results are representative of three independent experiments, each done in triplicate. Bars represent mean ± SD. (E) Immunoblot analysis of HCT15, SNUC5, COLO205, and HRT18 cell lines treated with 0.25 or 0.5 µ m MK‐1775 (n = 2). (F) GDSC2 analysis of indicated PI3K or AKT inhibitors according to ARID1A mutation status. Data are presented as scatter plots. Mean values are indicated by red lines. * p < 0.05, ** p < 0.01 (two‐tailed Wilcoxon rank‐sum test).

Journal: Advanced Science

Article Title: Targeting WEE1 in ARID1A/TP53 Concurrent Mutant Colorectal Cancer by Exploiting R‐Loop Accumulation and DNA Repair Deficiencies

doi: 10.1002/advs.202512074

Figure Lengend Snippet: CRISPR screening revealed that co‐targeting WEE1 and AKT as a potential combination strategy in ARID1A/TP53 concurrent mutant CRC. A) Schematic description of the CRISPR knockout kinome library screen. (B) Results of CRISPR screen in SNUC5 cells. Scatter plots present the Z‐scores of average log 2 (fold change) for MK‐1775 vs DMSO ( x ‐axis) and for ZN‐c3 vs DMSO ( y ‐axis). (C) Immunoblot analysis confirming AKT2 knockout in SNUC5 cells (n=2). (D) Dose‐response curves of SNUC5 cells, treated with MK‐1775 (left) or ZN‐c3 (right) for 5 days. The results are representative of three independent experiments, each done in triplicate. Bars represent mean ± SD. (E) Immunoblot analysis of HCT15, SNUC5, COLO205, and HRT18 cell lines treated with 0.25 or 0.5 µ m MK‐1775 (n = 2). (F) GDSC2 analysis of indicated PI3K or AKT inhibitors according to ARID1A mutation status. Data are presented as scatter plots. Mean values are indicated by red lines. * p < 0.05, ** p < 0.01 (two‐tailed Wilcoxon rank‐sum test).

Article Snippet: Human kinome CRISPR knockout library (Brunello) was obtained from Addgene (#73179) and was packaged into lentivirus.

Techniques: CRISPR, Mutagenesis, Knock-Out, Western Blot, Two Tailed Test