Abderrazzaq, H.; Singh, M.; Babb, L.; Bergquist, T.; Brenner, S. E.; Pejaver, V.; O'Donnell-Luria, A.; Radivojac, P.; ClinGen Computational Working Group, ; ClinGen Variant Classification Working Group,

Insertions and deletions (indels) represent a substantial source of genetic variation in humans and are associated with a diverse array of functional consequences. Despite their prevalence and clinical importance, indels, particularly short in-frame indels, remain critically understudied compared to single nucleotide variants and are challenging to interpret clinically. While many computational predictors for missense variants have been rigorously evaluated and calibrated for clinical use, the clinical utility of tools for in-frame indels remains uncertain. To address this gap, we have calibrated in-frame indel prediction tools for clinical variant classification. We constructed a high-confidence dataset of in-frame indel variants ([≤] 50bp) from clinical and population databases and estimated the prior probability of pathogenicity of a rare in-frame indel observed in a disease-associated gene, and of an insertion and deletion separately. Using a previously developed statistical framework based on local posterior probabilities, we then established score thresholds for eight computational tools, corresponding to distinct evidence levels for pathogenic and benign classification according to ACMG/AMP guidelines. All in-frame indel predictors evaluated here reached multiple evidence levels of pathogenicity and/or benignity, demonstrating measurable clinical value. However, these models consistently exhibited lower performance levels compared to missense predictors, highlighting the need for improved computational approaches for indel classification.