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. 2019 Mar 25;8:e43333. doi: 10.7554/eLife.43333

Figure 3. CRISPR-Cas9-mediated knock-out of WRN confirms the selective dependency of MSI-H CRC models on WRN.

(A) Schematic representation of CRISPR-Cas9 depletion assays. Cas9 expressing cells were transduced with a lentivirus encoding GFP and sgRNAs. The percentage of GFP-positive cells was determined over time by flow cytometry. (B) Cas9 expressing MSS or MSI-H CRC cells were transduced with a lentivirus encoding GFP and sgRNAs targeting multiple domains in WRN as indicated. The percentage of GFP-positive cells was determined 14 days post-transduction and normalized to the fraction of GFP-positive cells at the first measurement. Depletion ratios are shown relative to the positive control RPA3 (n = 1 experimental replicate). Domains are annotated according to PFAM entry Q14191. RQC, RecQ helicase family DNA-binding domain; HRDC, Helicase and RNase D C-terminal, HTH, helix-turn-helix motif.

Figure 3.

Figure 3—figure supplement 1. CRISPR-Cas9-mediated knock-out of WRN confirms the selective dependency of MSI-H CRC models on WRN.

Figure 3—figure supplement 1.

Cas9-GFP expressing MSS or MSI-H CRC cells were transduced with a lentivirus encoding GFP and sgRNAs targeting multiple domains in WRN as indicated. The percentage of GFP-positive cells was determined 14 days post-transduction and normalized to the fraction of GFP-positive cells at the first measurement. Depletion ratios are shown relative to the pan-essential positive control RPA3 (n = 1 experimental replicate). Domains are annotated according to PFAM entry Q14191. RQC, RecQ helicase family DNA-binding domain; HRDC, Helicase and RNase D C-terminal, HTH, helix-turn-helix motif.