Prime editing for the investigation of aberrant splicing defect associated with a pathogenic PRPH2 variant
Summary
The human Peripherin 2 (PRPH2) gene, essential for the structure and function of photoreceptor outer segments, is implicated in a range of inherited retinal diseases (IRDs). This study focuses on the pathogenic c.828+1G>A PRPH2 splice site variant. We employed prime editing (PE) technology to install and correct this variant in human induced pluripotent stem cells (hiPSCs). We developed an all-in-one PE construct, featuring a GFP reporter to facilitate the identification of successfully edited clones. The resulting heterozygous and homozygous hiPSC clones exhibited no detectable off-target mutations or karyotype abnormalities. Crucially, we found in hiPSCs and DD50/DD100 precursor hiPSC-derived retinal organoids that the c.828+1G>A PRPH2 mutation leads to activation of a cryptic splice site and intron retention, forming a mutant transcript. Importantly, correction of the c.828+1G>A PRPH2 mutation in the homozygous hiPSC clone resulted in the restoration of the canonical PRPH2 transcript and a reduction of the mutant transcript. Our findings highlight the potential of PE as a precise and safe method for installing and correcting pathogenic PRPH2 mutations in hiPSCs, paving the way for future genotype-phenotype studies and therapy development for PRPH2-mediated IRDs. © 2025 The Author(s).
| Authors | Lopes da Costa B, Helms KM, Theodore K, Tsai YT, Caruso SM, Liu S, Lima de Carvalho JR, Nolan ND, Tahir S, Makinson CD, Tsang SH, Quinn PMJ |
|---|---|
| Journal | Molecular therapy. Nucleic acids |
| Publication Date | 2025 Dec 9;36(4):102740 |
| PubMed | 41210588 |
| PubMed Central | PMC12594906 |
| DOI | 10.1016/j.omtn.2025.102740 |