Priya M

Does the abstract specify which rps12 intron is affected, so the reported defect can be mapped to a particular splice junction?

Given your report that patient samples were excluded, my same-sample check should refer specifically to construct-derived MYH9 RNA in the minigene assay. Patient-tissue junction measurements fall outside that study scope.

Did the matched wild-type minigene retain exon 29, or did it also produce the skipped product?

Separate wild-type and pseudoexon measurements don’t establish assay specificity. The primer and probe positions, or the wild-type amplicon sequence, are needed to tell whether the signal is restricted to the exon 44 to exon 45 junction.

The 5 of 18 read count makes presence of the specified aberrant junction interpretable, but leaves an isoform ratio poorly constrained. Was the canonical NM_001278116.2 junction detected in the same urine-derived-cell sample and in matched controls, with both junction assays inside a validated quantitative range?

Before treating an aberrant:canonical junction ratio as quantitative evidence, both junction assays should be shown to remain within their validated measurement range in every sample and matched comparator. A dilution series or equivalent linearity check would be decisive; otherwise, low canonical signal from limited L1CAM expression could inflate the apparent splice shift.

A detectable L1CAM transcript does not by itself validate an aberrant/normal isoform ratio. I would retain that ratio as quantitative RNA evidence only if both junction assays remained within their validated quantitative range in every case and comparator sample; otherwise, report the splice products qualitatively and separate that observation from any estimate of relative abundance.

“Detectable” needs an assay-level threshold. For the L1CAM RNA result, the key check is whether the canonical and aberrant junction measurements remain within the assay’s quantitative range in each urine-derived-cell sample, with the unaffected comparator processed in the same batch. Otherwise, an aberrant/normal ratio may be driven by a near-floor canonical signal rather than a stable isoform shift.