Resolving the GBA gene with Oxford Nanopore technology
About Marco Toffoli
Dr. Marco Toffoli MD is a clinical neurologist with an interest in movement disorders. He is a Ph.D. student at the Institute of Neurology, University College London. His main area of research involves the genetics of Parkinson's, and in particular the role of the GBA gene in the pathogenesis of the disease.
Variants in the GBA gene cause Gaucher disease and represent a major risk factor for Parkinson's disease. Sequencing of GBA is challenging because a highly homologous pseudogene causes a high frequency of complex recombinations. Our group recently described a novel approach for sequencing of GBA with MinION and PCR enrichment. One limitation was that the PCR misses reciprocal fusion and duplication recombinants. We overcame this by including two alternative PCRs to detect recombinants and refined the bioinformatics, now including Guppy, NGMLR, Clair and Sniffles. We screened over 300 individuals, detected several mutations, and verified the presence of common intronic haplotypes.