Fig. 4.

Crossing scheme for distinguishing maternal and paternal kmo allele editing. a Homozygous, white-eye female or male mosquitoes (open white symbols) with a deletion on the kmo gene exon 5 at nucleotide (NT) 460 were outcrossed to wild-type mosquitoes (black symbol) to identify the effects of Cas9 activity in single chromosomes (paternal or maternal). Wild type intercrosses were used to test for simultaneous gene editing in the paternal and maternal chromosomes. An “x” on the illustrated chromosome indicates mutation at that site. b Genotype and phenotype resulting from the G₀ crosses in the absence of Cas9 activity; there is no mutation on the 519 site, the progeny are either heterozygous for mutation at 460 or wild-type homozygous, and therefore the phenotype of individuals is wild type. c Genotype and phenotype resulting from the G₀ crosses in the presence of Cas9 activity at the 519 site. Progeny from intercrosses are white or mosaic eyes. Monoallelic activity at the 519 site is undetectable in G₀ progeny from the G₋₁ wild-type intercross, but biallelic activity is detectable as white eyes or mosaics. Further crossing of G₀ can lead to detection of monoallelic activity