The molecular complexity of COL2A1 splicing variants and their significance in phenotype severity

Pathogenic single nucleotide variants (SNVs) found in the COL2A1 gene are associated with a broad range of skeletal dysplasias due to their impact on the structure and function of the Col2a1 protein. However, the molecular mechanisms of some nucleotide variants detected during diagnostic testing remain unclear. The interpretation of missense and splicing variants caused by SNVs poses a significant challenge for clinicians. In this work, we analyzed 22 splicing variants in the COL2A1 gene which have been found in patients with COL2A1-associated skeletal dysplasias. Using a minigene system, we investigated the impact of these SNVs on splicing and gained insights into their molecular mechanisms and genotype-phenotype correlations for each patient. The results of our study are very useful for improving the accuracy of diagnosis and the management of patients with skeletal dysplasias caused by SNVs in the COL2A1 gene. •Establishing a molecularly confirmed diagnosis to patients with splice variants in the COL2A1 gene poses a challenge.•Functional analysis is essential procedure for pathogenicity evaluation and identification of splicing changes mechanisms.•We successfully analyzed 22 splicing variants in COL2A1. The obtained data allows to clarify the molecular mechanisms.