The Contribution of Non-Canonical Splice Sites Variants in Parkinson's Disease-Associated Genes to RNA Splicing

Parkinson's disease (PD) is the second most frequent neurodegenerative disorder. Non-canonical splice-site splicing-disrupt variants significantly contribute to neurological disorders. However, until recently, there was a lack of comprehensive evaluation of splicing-disrupt variants in PD-associated genes to PD. This study systematically assessed the contribution of non-canonical splice-site splicing-disrupt variants to PD. Whole-exome sequencing data of 1676 PD patients were analyzed using predictive tools SpliceAI and SPIDES. Twelve novel putative splicing-disrupt variants were predicted to affect precursor messenger RNA (pre-mRNA) splicing. Nine variants were verified by minigene splicing assay to determine whether they affected pre-mRNA splicing, and RT-PCR on the patients’ blood RNA validated the remaining three variants. Only a novel exon variant, GIGYF2 (NM_001103146: c.1079A>G) caused aberrant splicing, the remaining 11 variations were proved not to affect splicing. Collectively, we are the first to indicate the rare contribution of non-canonical splice-site variants in PD-associated genes to splicing in a large cohort.Funding Information: This study was funded by the National Key R&D Program of China (2021YFC2502100) and Hunan Youth Science and Technology Innovation Talent Project (2020RC3060) awarded to JC and Natural Science Foundation of Hunan province in China (2021JJ31070) to BL.Declaration of Interests: The authors declare no conflict of interest.Ethics Approval Statement: All 1676 patients with PD provided written informed consent for this genetic research, which was approved by the Oversight Committee and the Institutional Review Board of the Xiangya Hospital of Central South University.