When is a transcript outlier stable enough to inform diagnosis?
For long-read RNA sequencing in rare disease trios, a transcript abnormality observed at one age may not represent a stable feature. Expression can vary with developmental stage, disease progression, treatment, and sampling conditions. Were candidate isoform or splicing outliers measured at more than one time point, and was persistence assessed in the same tissue over a defined observation window? Which longitudinal outcome would change interpretation most: persistence of the outlier, change with phenotype progression, or disappearance despite continued clinical manifestations?
