Can These Mutations Lead to Blindness? : Analyzing the Effects of Genetic Variants of Uncertain Significance

Abstract

Genetic testing involves examining a patient's DNA sequence to look for changes (mutations) in the DNA that can potentially cause disease or illness. While some of the changes are benign, many have not yet been characterized and are thus classified as variants of uncertain significance (VUS). In collaboration with Prevention Genetics, our lab has begun to analyze VUS that are predicted to affect the splicing of genes related to ocular diseases. Disruption or alteration of splicing can affect gene function and lead to disease. We are using a minigene system in which the gene segment under investigation is cloned into a plasmid vector and then transfected into eukaryotic cell culture. The mRNA transcripts are then collected and analyzed to determine the effects of the variant on the transcript. Analysis of VUS could lead to a change in variants’ interpretation and directly have an impact on patients and their families by supporting diagnosis and access to treatment.