The PAX6 is essential for ocular morphogenesis and is known to be highly sensitive to changes in gene expression, where neither over- nor under-expression ensures normal ocular development. Two unrelated probands with classical aniridia who were previously considered “PAX6-negative”, were studied by whole-genome sequencing. Through the use of multiple in silico deep learning-based algorithms, we identified two novel putative causal mutations, c.-133_-132del in the 5′ untranslated region (5′-UTR) and c.-52 + 5G>A in an intron upstream of the PAX6 gene. The luciferase activity was significantly increased and VAX2 binding was disrupted with the former 5′-UTR variant compared with wild-type sequence, which resulted in a striking overexpression of PAX6. The minigene assay showed that the c.-52 + 5G>A mutation caused defective splicing, which resulted in the formation of truncated transcripts.

该PAX6是用于眼部形态必不可少的，被称为是对基因表达的变化，其中既不过高，也不表达不足确保正常眼发育高度敏感。通过全基因组测序研究了两个以前被认为是“ PAX6阴性”的经典无虹膜病先证者。通过使用多种基于计算机深度学习的算法，我们确定了两个新的推定因果突变，即5'非翻译区（5'-UTR）中的c.-133_-132del和一个5'-UTR中的c.-52 + 5G> A。PAX6上游的内含子基因。与野生型序列相比，萤光素酶活性显着增加，而前5'-UTR变体破坏了VAX2的结合，从而导致PAX6明显过表达。小基因检测表明，c.-52 + 5G> A突变引起有缺陷的剪接，从而导致截短的转录本的形成。