Targeting cryptic splicing variants in SDCCAG8, CEP290, and ABCA4: A translational workflow from diagnosis to ASO-based therapy
Year: 2026
Type: article
Abstract: A definitive molecular diagnosis is essential for the clinical management of rare genetic diseases (RGDs), guiding prognosis, genetic counseling, and therapeutic development. RNA-based therapeutics have emerged as promising strategies for RGDs owing to their ability to modulate gene expression and splicing. Here, we establish an integrated and scalable pipeline from variant discovery to RNA-based therapy in inherited retinal diseases (IRDs). Genome sequencing identified three cryptic variants in known IRD genes. Two novel deep intronic variants in SDCCAG8 were characterized by RT-PCR to induce pseudoexon inclusion. In addition, a previously described deep intronic variant in CEP290 was detected and reproduced on patient-derived fibroblasts, confirming aberrant splicing. A third family carrying the recurrent ABCA4 c.768G>T silent variant was included to corroborate the translational strength of our pipeline and to provide an additional model for RNA-based therapeutic correction. We designed and tested 16 splice-switching antisense oligonucleotides (ssASOs) in patient-derived fibroblasts, HeLa cells transfected with mutant minigenes, and, when available, human induced pluripotent stem cell (hiPSC)-derived retinal organoids. For each variant, at least one ssASO successfully improved normal splicing and increased gene expression in patient-derived models. Overall, this study validates an innovative end-to-end translational workflow and highlights how molecular diagnosis can directly drive the development of personalized RNA-based therapies.
Authors: Mariia Zadorozhna, Simona Rossi, Erika Tenderini, Ilaria Brentari, Anna Basile, Giulia Pezzin, Rebecca Venturini, Federica Isidori, Elena Luppi, Emanuele Panza, Fabio Sirchia, Elena Rossi, Mariateresa Zanobio, Marianthi Karali, Tommaso Pippucci, Giulio Piluso, Michela Alessandra Denti, Marco Seri, Sandro Banfi, Ivana Trapani, Elisa Giorgio
Language: English
Subfield: Ophthalmology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $3,900
OpenAlex ID: W7221777672