To improve the interpretation of genetic variants and support diagnosis in patients with rare diseases using a targeted long-read RNA sequencing approach.
Key Findings:
The approach identified all previously reported pathogenic variants in patients with known genetic diagnoses and provided additional insights into RNA processing.
Long-read sequencing enabled analysis across larger DNA regions, facilitating the determination of whether variants occur on the same or different alleles.
Variants affecting splice sites led to a wider range of RNA changes than expected, with complex effects observed in over half of the splice site variants examined.
The method identified disease-causing variants in patients who had not previously received a genetic diagnosis.
Interpretation:
The targeted long-read RNA sequencing approach enhances the understanding of genetic variants' effects on gene expression and splicing, potentially leading to better diagnostic outcomes for patients with rare diseases.
Limitations:
The method is limited to the genes included in the panel and may not detect variants in genes outside the selected set.
Further work is needed to standardize workflows and integrate the approach into existing diagnostic pathways before clinical implementation.
Conclusion:
The STRIPE method shows promise in improving genetic diagnosis for rare diseases, but further validation and integration into clinical practice are necessary.
