Tilch E, Schormair B, Zhao C, et al. - Given that restless legs syndrome is a frequent neurological disorder with a substantial burden on individual well-being and public health, researchers conducted the study for identifying restless legs syndrome genes by mutational load analysis. In 4,649 individuals and 4,982 controls by next-generation sequencing, 84 candidate genes were analyzed using molecular inversion probes that primarily targeted coding regions. Through one approach, 14 genes were highly significant and confirmed by the other with Bonferroni-corrected significance to display a differential burden of low-frequency and unusual variants in restless legs syndrome. Nine of these (AAGAB, ATP2C1, CNTN4, COL6A6, CRBN, GLO1, NTNG1, STEAP4, VAV3) resided in the vicinity of known restless legs syndrome loci, while 5 (BBS7, CADM1, CREB5, NRG3, SUN1) have not previously been linked to restless legs syndrome. The burden test and binomial performance deviation analysis also converged significantly within these genes in fine-mapping of potentially causative domains. The differential burden with intragenic low-frequency variants shows putatively causative genes in restless legs syndrome.