WRN rescue experiment.
SW620 and SW48 cells (2 × 10^5 cells) were transfected by nucleofection (Lonza 4D Nucleofector Unit X) with Cas9–sgRNA ribonucleoproteins (RNP) targeting human MAVS (used as a non-essential knockout control) or WRN, together with overexpression of 200 ng pmGFP control or 200 ng mouse Wrn cDNA (Origene, MR226496). From each sample after nucleofection, 5,000 cells were seeded in a 96-well plate and allowed to grow for 5 days, after which cells were collected for either CellTiter-Glo assay or western blot analysis. CellTiter-Glo data were read on an Envision Multiplate Reader and data analysis was performed using GraphPad Prism 7 software. Student’s t-test was performed using the multiple t-test module in Prism 7. The sgRNA sequences that were used are listed in Supplementary Table 10