The allele frequency of the c.563A>G (p.Lys188Arg) variant in GJB2 is 0.006% (1/15428) of European chromosomes by gnomAD, which is a low enough frequency to apply PM2 based on the thresholds defined by the ClinGen Hearing Loss Expert Panel for autosomal recessive hearing loss (PM2). This variant has been detected in four probands patients with hearing loss (PMID 17666888, University of Minnesota internal data, GeneDx internal data). For two of those probands, a pathogenic was observed in trans, and in one individual a second pathogenic variant was observed but phase was not determined (PM3_Strong; PMID 17666888, University of Minnesota internal data, GeneDx internal data). The REVEL computational prediction analysis tool produced a score of 0.9, which is above the threshold necessary to apply PP3; however, this information is not predictive of pathogenicity on its own. In summary, this variant meets criteria to be classified as likely pathogenic for autosomal recessive hearing loss based on the ACMG/AMP criteria applied as specified by the Hearing Loss Expert Panel (PM2, PM3_Strong, PP3).