The nanos^dintegral gene drive enables population modification of the malaria vector Anopheles gambiae

The modification of mosquito populations at scale through CRISPR-Cas9-mediated homing gene drives is a promising route for malaria vector control. Integral gene drives (IGDs) are designed to utilize the regulatory sequences of endogenous genes to express only the minimal set of components required for gene drive. In this study, we describe the creation and characterization of the nanos^dIGD targeting and inserted into thenanosgene of the malaria vectorAnopheles gambiaeand show that it achieves high rates of gene drive (97.7% in females, 99.0% in males). We find that homozygous nanos^dfemales but not males show impaired fecundity, and a variable loss of ovary phenotype. Transcriptomic analysis of ovaries points to decreased transcript levels of thenanosgene when harbouring Cas9. As a minimal genetic modification, nanos^ddoes not induce widespread transcriptomic perturbations, and its susceptibility toPlasmodiumspp. and O’nyong nyong virus infection remains similar to wild-type mosquitoes. Importantly, we find that nanos^dpropagates efficiently in caged mosquito populations and is maintained as a source of Cas9 after the emergence of drive resistant alleles whilst also mobilising a non-autonomous antiparasitic effector modification. The nanos^dgene drive shows promise as a genetic tool for malaria vector control via population modification, and we outline steps towards its further optimization.