Tau Protein pp 307-324 | Cite as

Finding MAPT Mutations in Frontotemporal Dementia and Other Tauopathies

  • Carol Dobson-Stone
  • John B. J. KwokEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1523)


Sanger sequencing is a classic technique in molecular genetics to detect single nucleotide DNA variants in genomic DNA. Here we describe the detection of MAPT mutations by polymerase chain reaction amplification of patient genomic DNA followed by bidirectional Sanger sequencing. Exon trapping is a technique whereby genomic DNA covering the exon of interest and flanking intronic sequence is cloned into the intron of an expression vector and transfected into human cell lines. RNA is extracted and splicing products are examined by reverse-transcriptase PCR and agarose gel electrophoresis. We outline the application of this technique to assess the effect of novel DNA variants on the splicing efficiency of MAPT exon 10, a common mechanism of disease for pathogenic MAPT mutations.

Key words

Polymerase chain reaction Sanger sequencing Heterozygote detection Exon trapping pSPL3 


  1. 1.
    Hutton M, Lendon CL, Rizzu P et al (1998) Association of missense and 5′-splice-site mutations in tau with the inherited dementia FTDP-17. Nature 393:702–705CrossRefPubMedGoogle Scholar
  2. 2.
    Ghetti B, Oblak AL, Boeve BF et al (2015) Invited review: frontotemporal dementia caused by microtubule-associated protein tau gene (MAPT) mutations: a chameleon for neuropathology and neuroimaging. Neuropathol Appl Neurobiol 41:24–46CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Josephs KA, Hodges JR, Snowden JS et al (2011) Neuropathological background of phenotypical variability in frontotemporal dementia. Acta Neuropathol 122:137–153CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Kertesz A (2003) Pick’s complex and FTDP-17. Mov Disord 18:S57–S62CrossRefPubMedGoogle Scholar
  5. 5.
    Goedert M, Spillantini MG, Jakes R et al (1989) Multiple isoforms of human microtubule-associated protein tau: sequences and localization in neurofibrillary tangles of Alzheimer’s disease. Neuron 3:519–526CrossRefPubMedGoogle Scholar
  6. 6.
    Goedert M, Jakes R (1990) Expression of separate isoforms of human tau protein: correlation with the tau pattern in brain and effects on tubulin polymerization. EMBO J 9:4225–4230PubMedPubMedCentralGoogle Scholar
  7. 7.
    Hong M, Zhukareva V, Vogelsberg-Ragaglia V et al (1998) Mutation-specific functional impairments in distinct tau isoforms of hereditary FTDP-17. Science 282:1914–1917CrossRefPubMedGoogle Scholar
  8. 8.
    D’Souza I, Poorkaj P, Hong M et al (1999) Missense and silent tau gene mutations cause frontotemporal dementia with parkinsonism-chromosome 17 type, by affecting multiple alternative RNA splicing regulatory elements. Proc Natl Acad Sci U S A 96:5598–5603CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Rabbani B, Tekin M, Mahdieh N (2014) The promise of whole-exome sequencing in medical genetics. J Hum Genet 59:5–15CrossRefPubMedGoogle Scholar
  10. 10.
    Apel TW, Scherer A, Adachi T et al (1995) The ribose 5-phosphate isomerase-encoding gene is located immediately downstream from that encoding murine immunoglobulin kappa. Gene 156:191–197CrossRefPubMedGoogle Scholar
  11. 11.
    Buckler AJ, Chang DD, Graw SL et al (1991) Exon amplification: a strategy to isolate mammalian genes based on RNA splicing. Proc Natl Acad Sci U S A 88:4005–4009CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Stanford PM, Shepherd CE, Halliday GM et al (2003) Mutations in the tau gene that cause an increase in three repeat tau and frontotemporal dementia. Brain 126:814–826CrossRefPubMedGoogle Scholar
  13. 13.
    Pickering-Brown SM, Baker M, Gass J et al (2006) Mutations in progranulin explain atypical phenotypes with variants in MAPT. Brain 129:3124–3126CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Neuroscience Research AustraliaSydneyAustralia
  2. 2.School of Medical SciencesUniversity of New South WalesSydneyAustralia

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