NM_001754.5(RUNX1):c.668A>G (p.Glu223Gly) is a missense variant predicted to cause the substitution of glutamic acid by glycine at amino acid 223 (p.E223G). The highest population minor allele frequency in gnomAD v2 is 0.0001146 (13/113452 alleles) in the non-Finnish European population (PM2_Supporting, BS1, and BA1 are not met). This variant has also been observed in gnomAD v3/v4 in one homozygous individual of Finnish descent, and there are no known reports of affected patients with homozygous RUNX1 variants, plus RUNX1-null mice do not survive (BP2). The germline variant has been published in 2-3 children with B-ALL (PMID: 26580448; PMID: 34166225) and in a 22-year-old female with thrombocytopenia (low platelet count) and mild bleeding (menorrhagia), congenital aortic valve abnormality, and periventricular nodular heterotopia of the cerebrum but a negative family history of thrombocytopenia and hematologic malignancy (DOI: 10.1155/2023/4738660). However, the variant's presence in the general population (gnomAD) precludes the application of PS4 at any strength level. The computational predictor REVEL gives a score of 0.799, which is neither above nor below the thresholds predicting a damaging or benign impact on RUNX1 function. However, in vitro functional data indicate that the mutant protein exhibits normal transcriptional activity in a luciferase reporter assay using U937 cells (~105-115% of WT activity, Fig. 4), normal DNA-binding and β-subunit interaction in an EMSA assay using Cos7 cells (Fig. 2), appropriate nuclear localization in NIH3T3 cells, and normal ubiquitination (Fig. 5) (PMID: 23817177) (BS3). In summary, this variant meets the criteria to be classified as likely benign for autosomal dominant hereditary thrombocytopenia and hematologic cancer predisposition syndrome based on the ACMG/AMP criteria applied, as specified by the ClinGen Myeloid Malignancy VCEP: BP2 and BS3.