Figure 7. Recommendations for F0 knockout screens.

(A) Suggestion for the design of an F0 screen, based on a three-step process: (1) selection of gRNAs; (2) verification that all gRNAs are mutagenic; (3) phenotyping. During round 1 (step 2), we recommend targeting a pigmentation gene such as slc24a5 to quantify success rate at injections before estimating minimum samples sizes and commencing the screen. (B) Minimum sample size to detect a phenotype at 0.8 statistical power and 0.05 significance level as more knockouts are present in the population of F0 larvae. Based on 10 simulations with 100 animals in each group (scrambled control, F0 knockout). Mean ± standard deviation across the 10 simulations. (left) Minimum sample sizes to detect the lack of response to mustard oil of the trpa1b knockouts. Dashed line indicates the real sample size of the experiment (n = 22, Figure 5A). (right) Minimum sample sizes to detect the lengthened circadian period of csnk1db knockouts. Dashed line indicates the real sample size of the experiment (n = 16, Figure 5B).