Multiplexed somatic CRISPR–Cas9 genome editing reveals that loss of Egr1 has the most pronounced and consistent effect on tumor initiation, growth, and progression in Kras-driven lung cancer (IMAGE)
Caption
Multiplexed somatic CRISPR–Cas9 genome editing reveals that loss of Egr1 has the most pronounced and consistent effect on tumor initiation, growth, and progression in Kras-driven lung cancer.
A) Individual tumor sizes ordered by mean effect of targeted gene knockout in KC mice at 6 weeks and 20 weeks after tumor initiation. The number of mice in each cohort is as indicated. Each dot represents a tumor. The area of each dot is proportional to the number of cancer cells in each tumor.
(B) Mean effect of gene knockout of each sgRNA in KC mice at 6 weeks after tumor initiation. LN mean is a maximum likelihood estimator (MLE) of the mean tumor size under a log-normal sampling distribution. Each gene in this study was targeted by two high-specificity sgRNAs, imparting consistent growth effects (Pearson's r = 0.86, P = 0.0029; see Materials and Methods).
(C) Quantile-Quantile (Q-Q) plot of tumor size distributions of the top 50th – 99.99th percentiles of the two Inert sgRNAs at both 6 and 20 weeks demonstrates reproducibility of size profiles.
(D) Analysis of relative tumor sizes in KRAS-driven tumors at 6 and 20 weeks after tumor initiation. Relative size of tumors for each targeted gene (sgTS) is divided by the respective sgInerts percentile and merged across replicate mice. Percentiles significantly different from sgInerts are in color (P < 0.05, two-sided bootstrap resampling). The darker the shade of color, the larger the percentile, as shown in the legend in gray scale. Error bars denote 95% confidence intervals also determined by bootstrap sampling.
(E) Summary of three distinct growth phenotype profiles. For each genotype, the number of tumors observed at six weeks after tumor initiation, alongside MLE of mean size at six weeks (tumor growth; see Materials and Methods), and the size of the largest tumors at 20 weeks (advanced progression) is depicted. All statistics are divided by their respective values for sgInerts. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 (two-sided bootstrap resampling).
Credit
Athar Khalil, Trang Dinh, Meaghan Parks, Rebecca C. Obeng, Berkley Gryder, Adam Kresak, Yuxiang Wang, Jeff Maltas, Madeline Bedrock, Xiangzhen Wei, Zachary Faber, Mira Rahm, Jacob Scott, Thomas LaFramboise, Zhenghe Wang, Christopher McFarl
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