2023 ACMG Annual Clinical Genetics Meeting Digital Edition: Poster Gallery-MINIGENE SPLICING ASSAY CONFIRMS the PATHOGENICITY of the NON-CANONICAL SPLICE SITE VARIANT c.11+5G>A in <em>RPE65</em>

MINIGENE SPLICING ASSAY CONFIRMS the PATHOGENICITY of the NON-CANONICAL SPLICE SITE VARIANT c.11+5G>A in RPE65

Pdf Summary

This study discusses a non-canonical splice site variant in the RPE65 gene, c.11+5G>A, which has been associated with retinopathy. The variant affects the splicing of the gene, resulting in reduced levels of mature mRNA. The prevalence of this variant is relatively high and it has been found in both homozygous and compound heterozygous states in multiple families. The variant has also been reported in individuals with a founder mutation in trans. The minigene splicing assay confirms that the variant affects splicing, with the variant construct showing significantly lower levels of mature mRNA compared to the reference construct. The variant is classified as pathogenic according to the ACMG/AMP guidelines based on evidence codes PS3, PS4, PM2_Supporting, PM3, and PP1. The study highlights the importance of experimental verification in accurately interpreting genetic variants, especially for disorders that have therapeutic interventions. The results of this study suggest a potential novel splicing mechanism leading to disease and provide valuable information for diagnostic purposes and therapeutic interventions for RPE65-related retinopathies.

Asset Subtitle

Presenting Author - Madhulatha Pantrangi, PhD; Co-Author - Bin Guan, PhD, FACMG; Co-Author - Kristy Lee, MS, CGC; Co-Author - Nia Moore, BS; Co-Author - William Hankey, PhD; Co-Author - Robert B. Hufnagel, MD, PhD;

Meta Tag

Databases

Eye disorders

Genetic Testing

Methodology

Population Genetics

Sequencing

Variant Detection

Co-Author Bin Guan, PhD, FACMG

Co-Author Kristy Lee, MS, CGC

Co-Author Nia Moore, BS

Co-Author William Hankey, PhD

Co-Author Robert B. Hufnagel, MD, PhD

Presenting Author Madhulatha Pantrangi, PhD

Keywords

non-canonical splice site variant

RPE65 gene

retinopathy

mature mRNA

prevalence

homozygous

compound heterozygous

founder mutation

minigene splicing assay

pathogenic variant