Related Experiment Video
Updated: Jul 2, 2026
09:34
Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease
Published on: April 4, 2018
Variant-specific RNA testing resolves variants of uncertain significance in exome testing
Audrey K O'Neill1, Grace E VanNoy2, Brooklynn Gasser1
1Ambry Genetics, 1 Enterprise Drive, Aliso Viejo, California, 92656, USA.
BMC Genomics
|July 1, 2026
Summary
Resolving variants of uncertain significance (VUS) in genetic testing is crucial. Targeted RNA testing effectively reclassified VUS, leading to accurate diagnoses in all cases studied.
Area of Science:
- Genetics and Genomics
- Molecular Biology
- Clinical Diagnostics
Background:
- Variants of uncertain significance (VUS) pose challenges in genetic diagnostics.
- VUS affecting splicing are particularly difficult to interpret.
- Exome sequencing (ES) is a powerful tool, but VUS interpretation remains a hurdle.
Purpose of the Study:
- To demonstrate the utility of variant-specific RNA testing for VUS resolution.
- To reclassify VUS with limited direct evidence.
- To improve molecular diagnoses in patients undergoing exome testing.
Main Methods:
- Exome sequencing (ES) followed by variant-specific RNA testing.
- Analysis of VUS in four distinct clinical cases.
- Assessment of splicing impact and protein sequence alterations.
Main Results:
- Four VUS were successfully reclassified to Likely Pathogenic.
- Aberrant splicing was confirmed for all tested variants.
- Positive molecular diagnoses were achieved in all four cases.
- Variants identified in ATP6V0A2, SLC20A2, CSF1R, and COL1A1.
Conclusions:
- Targeted RNA testing is highly effective for reclassifying VUS.
- This approach resolves VUS with limited direct evidence.
- RNA testing broadens the applicability of exome sequencing in clinical genetics.
Related Concept Videos
03:21RNA-seq
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while microarray-based...
02:24Ribosome Profiling
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
02:26Comparing Copy Number Variations and SNPs
Sequencing of the human genome has opened up several best-kept secrets of the genome. Scientists have identified thousands of genome variations that exist within a population. These variations can be a single nucleotide or a larger chromosomal variation.
Copy number variations or CNVs are the structural variations that cover more than 1kb of DNA sequence. The single nucleotide polymorphism (SNP), on the other hand, is a single nucleotide change or a point mutation that is found in more than 1%...
Copy number variations or CNVs are the structural variations that cover more than 1kb of DNA sequence. The single nucleotide polymorphism (SNP), on the other hand, is a single nucleotide change or a point mutation that is found in more than 1%...