The NM_000156.6:c.160G>C variant in GAMT is predicted to result in the substitution of alanine by proline at amino acid 54 (p.Ala54Pro). Three probands with clinical symptoms consistent with GAMT deficiency have been reported to be homozygous for this variant. One of these individuals had elevated urinary guanidinoacetate, and GAMT enzymatic activity <0.1 nmol/h/mg in lymphoblasts, with full GAMT gene sequencing performed (PMID: 15108290) (PP4_Strong). This individual harbored the variant in cis with two homozygous variants that have been classified as benign by the ClinGen CCDS VCEP, c.626C>T (p.Thr209Met) (ClinVar ID: 21068) and c.459+71G>A (aka c.460-31G>A, ClinVar ID: 21067). For the other two probands, neither biochemical nor MRS data were available (PMID: 38020815, 39006040); therefore, their data was not included for PM3 (total 0.5 points, PM3_Supporting). The variant is absent in gnomAD v4.1.0. (PM2_Supporting). The computational predictor REVEL gives a score of 0.894 which is in the range of 0.773-0.932, evidence that correlates with impact to GAMT function at the moderate level (PMID: 36413997) (PP3_Moderate). There is a ClinVar entry for this variant (Variation ID: 2446454). In summary, this variant meets the criteria to be classified as likely pathogenic for guanidinoacetate methyltransferase (GAMT) deficiency. GAMT-specific ACMG/AMP criteria applied, as specified by the ClinGen Cerebral Creatine Deficiencies Variant Curation Expert Panel (CCDS VCEP) (Specifications version 2.0.0): PP4_Strong, PP3_Moderate, PM2_Supporting, PM3_Supporting (Classification approved by the ClinGen Cerebral Creatine Deficiencies Variant Curation Expert Panel on October 8, 2025)