Fig. 1
Experimental parameters affecting Cas9-mediated mutant mouse line production. (a) Violin plot for founder rates from experiments with different methods of reagent delivery (CY, cytoplasmic injection; EP, electroporation; PN, pronuclear injection). Reported are the p-values (p) of pairwise comparisons using the Wilcoxon rank sum test, the number of genes (N), and corresponding medians in boxplots. (b) Violin plot for founder rates from experiments using two (2) or four (4) guide RNAs designed to produce deletion alleles. Reported is the p-value of pairwise comparison using the Wilcoxon rank sum test, the number of genes (N) and corresponding medians in boxplots. (c) Violin plot for founder rates from experiments with Cas9 guide RNAs designed to delete different sizes (in bp) of critical regions (genomic DNA). Each bin has ~ 640 unique gene deletion attempts. Reported is the p-value (p) of the overall comparison using the Kruskal–Wallis test. (d) Barplot showing the percentage of genes with GLT of the desired deletion allele after breeding one (1), two (2), three (3) or four or more (4 +) founders. Pairwise comparison of GLT rates using the Pearson chi-square test with Holm’s correction showed a significant difference only when breeding one founder was compared to breeding three or four founders (** p = 0.004 for each comparison, 1 vs. 3 and 1 vs. 4 +). No difference between 1 vs.2 or 3 vs. 4 + was observed. Unique gene attempts are the first attempt with GLT of the desired allele or the last of a set of unsuccessful attempts for each gene. See materials and methods for a complete description of data filtering. GLT, germline transmission.
