Fig 2. Establishment and maintenance of heritable loss-of-function mutations in Ae. aegypti flight genes.

(A) For each gene, the wild type (WT) and mutant sequence is shown. The PAM sites for each sgRNA used in the injection mix for the specified location are highlighted in red. Individuals containing each deletion were outcrossed through the indicated generation, at which point individuals heterozygous for the AeAct-4, myo-fem (B), and Aeflightin (C) mutations were intercrossed. For each cross, the ratio of each potential genotype expected is noted in parentheses, with the individuals with an expected flightless phenotype highlighted in red. For Aeflightin, phenotypic identification of all white-eyed pupae enables their removal before further phenotypic analysis based on flight (see Fig 4) is performed.