Our study identifies a homozygous missense variant (c.G5408A) in DNAH17, a functionally uncharacterized gene, from a consanguineous Pakistani family with three offspring suffering from asthenozoospermia (no MMAF-like phenotype) and provides genetic evidence that DNAH17 c.G5408A is pathogenic for asthenozoospermia using Dnah17M/M mice modeling the patients’ mutation. Here, DNAH17 is linked to Reduced sperm motility.