Diagnosis of mitochondrial disorders is still hampered by their phenotypic and genotypic heterogeneity. In many cases, exome sequencing, the state-of-the-art method for genetically diagnosing mitochondrial disease patients, does not allow direct identification of the disease-associated gene but rather results in a list of variants in candidate genes. Here, we present a method to validate the disease-causing variant based on functional complementation assays. First, cell lines expressing a wild-type cDNA of the candidate genes are generated by lentiviral infection of patient-derived fibroblasts. Next, oxidative phosphorylation is measured by the Seahorse XF analyzer to assess rescue efficiency.
CITATION STYLE
Kremer, L. S., & Prokisch, H. (2017). Identification of disease-causing mutations by functional complementation of patient-derived fibroblast cell lines. In Methods in Molecular Biology (Vol. 1567, pp. 391–406). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6824-4_24
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