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. Author manuscript; available in PMC: 2021 Nov 18.
Published in final edited form as: Cell Syst. 2020 Oct 19;11(5):523–535.e9. doi: 10.1016/j.cels.2020.09.009

Figure 3. The expression-fitness curves of essential genes in E. coli and B. subtilis can be studied using singly mismatched sgRNAs.

Figure 3.

(A) Schematic of the fitness experiment design. Each library was measured in 4-6 replicates and averaged. (B) Distribution of per sgRNA locus (solid lines) and per gene (dashed lines) correlations (Pearson r) for sgRNAs targeting genes in E. coli (orange) and B. subtilis (blue). (C-D) The fitness effects of all fully complementary sgRNA targeting essential genes in E. coli (C) and B. subtilis (D) showing that the identity of the targeted gene is the driving factor in determining the fitness effect of an sgRNA. Genes are arranged in order of median fitness defect. Values represent the mean of 4-6 biological replicates. Individual values for each sgRNA and their standard deviations can be found in Table S3. The standard deviation of relative fitness of our 1,000 non-targeting control sgRNAs was 0.0825 in E. coli and 0.0444 in B. subtilis, indicating that even subtle growth defects of 5-10% can be accurately resolved.