A mutation-agnostic and allele-specific ASO strategy demonstrates potent functional rescue and retinal preservation in RHO-linked retinitis pigmentosa
- 1Institute for Ophthalmic Research, Centre for Ophthalmology, University of Tuebingen
Abstract
Autosomal dominant retinitis pigmentosa (adRP) caused by RHO mutations is a leading form of inherited retinal degeneration. Extensive allelic heterogeneity of RHO pathogenic variants limits the translational applicability of mutation-specific gene therapies. To address this, we developed SNARE (SNP-guided Silencing of Aberrant RHO Expression), a mutation-independent, allele-specific antisense oligonucleotide (ASO) strategy. SNARE selectively suppresses mutant RHO transcripts by targeting the common, benign c.-26A/G single-nucleotide polymorphism (SNP) as an allelic discriminator. Candidate gapmer ASOs were screened in engineered reporter lines and validated in patient-derived retinal organoids, identifying RHOligo-A as the lead c.-26A-targeting candidate. In vitro, RHOligo-A achieved robust, preferential knockdown of the target allele, improving RHO localization in retinal organoids, and demonstrated a favorable safety profile with minimal transcriptomic off-target effects and no detectable immunostimulatory activity. Subsequent validation in a novel, humanized RHOP347L/WT mouse model, achieved sustained c.-26A-linked allele-selective suppression, retinal structure preservation, and significantly restored visual function, upon a single intravitreal administration. These findings establish RHOligo-A and SNARE as a scalable, mutation-independent therapeutic platform with strong translational potential and substantial clinical reach for RHO-associated adRP.
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BibTeX
@misc{spaag2026mutationagnostic,
title = {{A mutation-agnostic and allele-specific ASO strategy demonstrates potent functional rescue and retinal preservation in RHO-linked retinitis pigmentosa}},
author = {S. Spaag and W.-H. Wu and J. Yun and T. Winogrodzki and A. S. Knudsen and M. Fuso and K. Stingl and G. Komissarov and A. Armento and B. Baumann and L. Kuehlewein and C. Ayuso and L. Fernandez-Caballero and R. Collin and Z. Corradi and S. Roosing and M. Kaltak and C. Lochmann and F. Radboudumc and S. Banfi and M. Karali and S. Bolz and F. Simonelli and K. Dave and S. Kohl and E. Zrenner and A. Demirkol and K. Achberger and B. Wissinger and S. H. Tsang and P. De Angeli},
howpublished = {bioRxiv},
year = {2026},
month = aug,
doi = {10.64898/2026.08.25.747013},
url = {https://www.biorxiv.org/content/10.64898/2026.08.25.747013v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - A mutation-agnostic and allele-specific ASO strategy demonstrates potent functional rescue and retinal preservation in RHO-linked retinitis pigmentosa
AU - S. Spaag
AU - W.-H. Wu
AU - J. Yun
AU - T. Winogrodzki
AU - A. S. Knudsen
AU - M. Fuso
AU - K. Stingl
AU - G. Komissarov
AU - A. Armento
AU - B. Baumann
AU - L. Kuehlewein
AU - C. Ayuso
AU - L. Fernandez-Caballero
AU - R. Collin
AU - Z. Corradi
AU - S. Roosing
AU - M. Kaltak
AU - C. Lochmann
AU - F. Radboudumc
AU - S. Banfi
AU - M. Karali
AU - S. Bolz
AU - F. Simonelli
AU - K. Dave
AU - S. Kohl
AU - E. Zrenner
AU - A. Demirkol
AU - K. Achberger
AU - B. Wissinger
AU - S. H. Tsang
AU - P. De Angeli
PY - 2026
DA - 2026/08/25
PB - bioRxiv
DO - 10.64898/2026.08.25.747013
UR - https://www.biorxiv.org/content/10.64898/2026.08.25.747013v1
AB - Autosomal dominant retinitis pigmentosa (adRP) caused by RHO mutations is a leading form of inherited retinal degeneration. Extensive allelic heterogeneity of RHO pathogenic variants limits the translational applicability of mutation-specific gene therapies. To address this, we developed SNARE (SNP-guided Silencing of Aberrant RHO Expression), a mutation-independent, allele-specific antisense oligonucleotide (ASO) strategy. SNARE selectively suppresses mutant RHO transcripts by targeting the common, benign c.-26A/G single-nucleotide polymorphism (SNP) as an allelic discriminator. Candidate gapmer ASOs were screened in engineered reporter lines and validated in patient-derived retinal organoids, identifying RHOligo-A as the lead c.-26A-targeting candidate. In vitro, RHOligo-A achieved robust, preferential knockdown of the target allele, improving RHO localization in retinal organoids, and demonstrated a favorable safety profile with minimal transcriptomic off-target effects and no detectable immunostimulatory activity. Subsequent validation in a novel, humanized RHOP347L/WT mouse model, achieved sustained c.-26A-linked allele-selective suppression, retinal structure preservation, and significantly restored visual function, upon a single intravitreal administration. These findings establish RHOligo-A and SNARE as a scalable, mutation-independent therapeutic platform with strong translational potential and substantial clinical reach for RHO-associated adRP.
ER -