Inherited retinal degenerations (IRDs) are at the focus of current genetic therapeutic advancements. For a genetic treatment such as gene therapy to be successful, an accurate genetic diagnostic is required. Genetic diagnostics relies on the assessment of the probability that a given DNA variant is pathogenic. Non-coding variants present a unique challenge for such assessments as compared to coding variants. For one, non-coding variants are present at much higher number in the genome than coding variants. In addition, our understanding of the rules that govern the non-coding regions of the genome is less complete than our understanding of the coding regions. Methods that allow for both the identification of candidate non-coding pathogenic variants and their functional validation may help overcome these caveats allowing for a greater number of patients to benefit from advancements in genetic therapeutics. We present here an unbiased approach combining whole genome sequencing (WGS) with patient-induced pluripotent stem cell (iPSC)-derived retinal organoids (ROs) transcriptome analysis. With this approach, we identified and functionally validated a novel pathogenic non-coding variant in a small family with a previously unresolved genetic diagnosis.

中文翻译：

---

RNA-seq 和全基因组测序相结合的方法，用于鉴定单个家族中的非编码致病变异。

遗传性视网膜变性（IRD）是当前基因治疗进展的焦点。为了使基因治疗（例如基因治疗）取得成功，需要准确的基因诊断。遗传诊断依赖于对给定 DNA 变异致病概率的评估。与编码变体相比，非编码变体对此类评估提出了独特的挑战。其一，基因组中非编码变体的数量远高于编码变体。此外，我们对基因组非编码区规则的理解不如对编码区的理解完整。允许识别候选非编码致病变异及其功能验证的方法可能有助于克服这些警告，使更多的患者受益于基因治疗的进步。我们在此提出一种将全基因组测序（WGS）与患者诱导多能干细胞（iPSC）衍生的视网膜类器官（RO）转录组分析相结合的无偏见方法。通过这种方法，我们在一个先前未解决基因诊断的小家族中发现了一种新的致病性非编码变异并进行了功能验证。