NM_001034853.2(RPGR):c.494G>T (p.Gly165Val) is a missense variant causing substitution of glycine by valine at amino acid 165. Another missense variant in the same codon, NM_001034853.2(RPGR):c.494G>A (p.Gly165Asp), has been classified as pathogenic for RPGR-related retinopathy by the ClinGen X-linked IRD VCEP, while no benign missense variants have been identified in this codon. The present variant has a higher Grantham’s distance (109) than the comparison variant (94), and SpliceAI has been used to confirm that neither variant has a predicted impact on RPGR splicing (PM5_Supporting). This variant is absent from gnomAD v4.1.0 (PM2_Supporting). This variant has been reported in at least 2 apparently unrelated probands meeting one of the PS4 requirements of a male with some functional vision impairment by age 30 years and/or decreased or absent electroretinogram responses, or a female with functional visual abnormality and documentation of a male relative affected with retinitis pigmentosa (PMID: 34745198, PMID: 31630094, PS4_Supporting). The computational predictor REVEL gives a score of 0.982, which is above the ClinGen X-linked IRD VCEP threshold of ≥ 0.932 and predicts a damaging effect on RPGR function (PP3_Strong). The splicing predictor SpliceAI gives a delta score of 0.06 for splice donor loss, which is lower than the threshold of >0.2 and does not strongly predict a disruption of splicing. Injection of human RPGR mRNA harboring the variant into zebrafish embryos rescued ciliary defects associated with rpgr silencing comparably to wild-type RPGR (PMID: 19815619). However, the BS3_Supporting code is not considered applicable for RPGR. In summary, this variant is classified as likely pathogenic for RPGR-related retinopathy based on the ClinGen X-linked Inherited Retinal Disease Expert Panel Specifications to the ACMG/AMP Variant Interpretation Guidelines for RPGR Version 1.0.0; PM2_Supporting, PP3_Strong, and PS4_Supporting.