NM_000329.3(RPE65):c.859G>T (p.Val287Phe) is both a missense variant, changing the valine at position 287 to phenylalanine, and a possible splicing variant, located at the first nucleotide of exon 9. SpliceAI's highest score is for Acceptor Loss, at 0.07, predicting a non-deleterious effect on splicing. The computational predictor REVEL has a score of 0.887, which is above the ClinGen LCA / eoRD VCEP threshold of ≥ 0.774 and predicts a damaging effect on RPE65 function (PP3_moderate). This variant is present in gnomAD v.4.1.1 at a GrpMax allele frequency of 0.00001426, with 4 alleles / 1613770 total alleles in the South Asian population, which is lower than the ClinGen LCA / eoRD VCEP PM2_Supporting threshold of <0.0002 (PM2_Supporting). At least one proband harboring this variant exhibits a phenotype including diagnosis of LCA (0.5 pts) and significant improvement after gene therapy treatment (8 pts), which together are highly specific for RPE65-related recessive retinopathy (8.5 points, PMID: 21911650 , PP4_Moderate). This variant has been reported in at least 2 unrelated probands with early-onset severe retinal dystrophy who were homozygous for the variant (1 point, PMIDs: 18722466, 21911650). This variant has also been reported in at least 1 proband with early-onset severe retinal dystrophy who was compound heterozygous with the NM_000329.3(RPE65):c.938A>G (p.His313Arg) variant confirmed in trans (1 point, VCEP member-provided data), which was previously classified pathogenic by the ClinGen LCA/eoRD VCEP (2 total points, PM3_Strong). In summary, this variant meets the criteria to be classified as Likely Pathogenic for RPE65-related recessive retinopathy based on the ACMG/AMP criteria applied, as specified by the ClinGen LCA/eoRD VCEP: PP3_Moderate, PM2_Supporting, PP4_Moderate, and PM3_Strong. (VCEP specifications version 1.0.0)