Detecting and Validating MAPT Mutations in Neurodegeneration Patients and Analysis of Exon Splicing Consequences

Methods Mol Biol. 2024:2754:411-433. doi: 10.1007/978-1-0716-3629-9_22.

Abstract

Mutation of MAPT has been observed in patients with parkinsonism, progressive supranuclear palsy, and corticobasal degeneration and is a significant cause of frontotemporal dementia. In this chapter, we discuss considerations for next-generation sequencing analysis to identify MAPT mutations in patient genomic DNA and describe the validation of these mutations by Sanger sequencing. One of the most common effects of MAPT mutations is differential splicing of exon 10, which leads to an imbalance in the proportion of 3-repeat and 4-repeat tau isoforms. We describe how to investigate the effect of novel DNA variants on the splicing efficiency of this exon in vitro using the exon-trapping technique, also known as the splicing reporter minigene assay.

Keywords: Exon trapping; Heterozygote detection; Long-read sequencing; Polymerase chain reaction; Sanger sequencingSanger sequencing; Splicing reporter minigene assay; Whole exome sequencingWhole exome sequencing (WES); Whole genome sequencingWhole genome sequencing (WGS); pSPL3.

MeSH terms

  • DNA
  • Exons
  • Frontotemporal Dementia* / genetics
  • Humans
  • Mutation
  • RNA Splicing
  • tau Proteins* / genetics

Substances

  • tau Proteins
  • DNA
  • MAPT protein, human