Genome sequencing reveals novel pathogenic deep-intronic PCDH15 variants, amenable to antisense oligonucleotide-based splice correction

Rodenburg, K.; Fenwick, L.; Pennings, R.; Haer-Wigman, L.; Ben-Yosef, T.; van Erp, F.; Reurink, J.; Gilissen, C.; van den Born, L. I.; Cremers, F. P. M.; Cohen, Y.; Yntema, H.; de Vrieze, E.; Kremer, H.; de Bruijn, S. E.; Collin, R. W. J.; Roosing, S.; van Wijk, E.

doi:10.64898/2026.08.20.746067

bioRxiv · 10.64898/2026.08.20.746067

Genome sequencing reveals novel pathogenic deep-intronic PCDH15 variants, amenable to antisense oligonucleotide-based splice correction

Rodenburg, K.·Fenwick, L.·Pennings, R.·Haer-Wigman, L.·Ben-Yosef, T.·van Erp, F.·Reurink, J.·Gilissen, C.·van den Born, L. I.·Cremers, F. P. M.·Cohen, Y.·Yntema, H.·de Vrieze, E.·Kremer, H.·de Bruijn, S. E.·Collin, R. W. J.·Roosing, S.·van Wijk, E.

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Abstract

Despite substantial advances in diagnostic testing, 10-15% of Usher syndrome patients remain without a genetic diagnosis, having significant implications for genetic counseling and potential future therapeutic interventions. In this study, genome sequencing data from probands clinically presenting with Usher syndrome were analyzed. Two novel deep-intronic variants were identified in PCDH15, c.3983+3635A>G and c.3123-1728A>G, in two independent patients. Both deep-intronic variants were classified as likely pathogenic and predicted to alter PCDH15 pre-mRNA splicing. Using a minigene splice assay and iPSC-derived photoreceptor precursor cells from patients, we confirmed that both variants lead to the inclusion of a pseudoexon in the PCDH15 transcript introducing a stop codon and subsequent premature termination of protein translation. We designed and evaluated antisense oligonucleotides (ASOs) with the purpose of redirecting aberrant pre-mRNA splicing caused by both deep-intronic variants. For both variants, designed ASOs were successful in restoring normal splicing patterns, highlighting their potential as a future therapeutic intervention strategy to halt the progression of retinitis pigmentosa caused by these novel variants. Overall, these findings contribute to the understanding of Usher syndrome caused by deep-intronic pathogenic variants in PCDH15 and describe for the first time the use of an ASO-mediated splice correction strategy for individuals diagnosed with these variants.

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Rodenburg, K., Fenwick, L., Pennings, R., Haer-Wigman, L., Ben-Yosef, T., van Erp, F., Reurink, J., Gilissen, C., van den Born, L. I., Cremers, F. P. M., Cohen, Y., Yntema, H., de Vrieze, E., Kremer, H., de Bruijn, S. E., Collin, R. W. J., Roosing, S., van Wijk, E.. 2026-08-24. Genome sequencing reveals novel pathogenic deep-intronic PCDH15 variants, amenable to antisense oligonucleotide-based splice correction. https://doi.org/10.64898/2026.08.20.746067

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