Detection of CGG repeats in Fragile X Syndrome using CRISPR Cas9 combined with single molecule sequencing
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Fragile X Syndrome is caused by CGG triplet-repeat expansions in the 5' untranslated region of the FMR1 gene. These alterations range from a pre-mutation state (55-200 repeats) to a full mutation allele (> 200 repeats). Triplet counting and allele methylation status are the two essential factors for diagnosing the disease. The project investigates the use of faster and cheaper nanopore sequencing technology (Oxford Nanopore), in combination with targeted editing by CRISPR Cas9 as an alternative technique for the simultaneous determination of triplet occurrences and DNA methylation status. This approach if proved to be effective could be generalised to detect many other dynamic mutations causing human diseases.