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Updated: Jun 9, 2026
08:53
A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
Published on: September 15, 2021
mRNA Sequencing to Identify Aberrant Splicing in X-linked Alport Syndrome
Dipti Rao1, Bartholomeus T van den Berge1,2, Anneke T Vulto-van Silfhout3
1Department of Nephrology, Radboud University Medical Center, Nijmegen, The Netherlands.
Kidney International Reports
|June 8, 2026
Summary
mRNA analysis is crucial for diagnosing X-linked Alport syndrome (XLAS) when standard genetic tests fail. It confirms variants of unknown significance and helps find deep-intronic mutations, improving genetic diagnosis for kidney disease.
Area of Science:
- Genetics
- Nephrology
- Molecular Biology
Background:
- X-linked Alport syndrome (XLAS) is a genetic kidney disease caused by COL4A5 gene variants.
- Routine genetic analysis misses pathogenic variants in 10-20% of XLAS patients.
Purpose of the Study:
- To evaluate the utility of mRNA analysis in diagnosing XLAS when conventional genetic testing is inconclusive.
- To identify novel pathogenic variants in patients with suspected XLAS.
Main Methods:
- mRNA analysis was performed on fibroblasts or urine-derived podocyte-lineage cells.
- Genomic DNA sequencing was guided by mRNA analysis findings.
- Variants of unknown significance (VUS) were assessed for pathogenicity using mRNA analysis.
Main Results:
- mRNA analysis confirmed pathogenicity in 2 patients with VUS.
- Novel deep-intronic pathogenic variants were identified in 7 out of 9 patients.
- Aberrant splicing was complete in 5 patients and partial in 4, correlating with kidney disease severity.
Conclusions:
- mRNA analysis is essential for confirming pathogenicity and identifying intronic variants in XLAS.
- This method aids genetic diagnosis in Alport syndrome (AS) cases where routine analysis is negative.
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