Functional Evaluation of Splice Variants Using a Minigene Strategy

Suspected splicing variants can be very challenging to evaluate more particularly when RNA samples are not available to perform RNA sequencing or when the gene of interest is expressed in specific tissues not easily reachable. One alternative strategy to study the potential splicing effect of a variant is to use the patient's DNA. Indeed, it is easily possible to amplify the region of interest and include it in the reporting system such as hybrid minigene, which could then be transfected in eukaryotic cells in order to estimate the impact of the mutated region in comparison to a wild-type version of the region. This strategy could help to evaluate the impact of genetic alterations on RNA splicing by using only patient's DNA.