Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus

Gillian Hamilton, Richard Killick, Jean-Charles Lambert, Philippe Amouyel, Minerva M Carrasquillo, V Shane Pankratz, Neill R Graff-Radford, Dennis W Dickson, Ronald C Petersen, Steven G Younkin, John F Powell, Richard Wade-Martins, Genetic and Environmental Risk for Alzheimer's Disease (GERAD1) Consortium, Patrick Kehoe

Research output: Contribution to journalArticle (Academic Journal)peer-review

4 Citations (Scopus)

Abstract

Nicastrin (NCSTN) is a component of the γ-secretase complex and therefore potentially a candidate risk gene for Alzheimer's disease. Here, we have developed a novel functional genomics methodology to express common locus haplotypes to assess functional differences. DNA recombination was used to engineer 5 bacterial artificial chromosomes (BACs) to each express a different haplotype of the NCSTN locus. Each NCSTN-BAC was delivered to knockout nicastrin (Ncstn(-/-)) cells and clonal NCSTN-BAC(+)/Ncstn(-/-) cell lines were created for functional analyses. We showed that all NCSTN-BAC haplotypes expressed nicastrin protein and rescued γ-secretase activity and amyloid beta (Aβ) production in NCSTN-BAC(+)/Ncstn(-/-) lines. We then showed that genetic variation at the NCSTN locus affected alternative splicing in human postmortem brain tissue. However, there was no robust functional difference between clonal cell lines rescued by each of the 5 different haplotypes. Finally, there was no statistically significant association of NCSTN with disease risk in the 4 cohorts. We therefore conclude that it is unlikely that common variation at the NCSTN locus is a risk factor for Alzheimer's disease.

Original languageEnglish
Pages (from-to)1848.e1-13
JournalNeurobiology of Aging
Volume33
Issue number8
DOIs
Publication statusPublished - Aug 2012

Bibliographical note

Copyright © 2012 Elsevier Inc. All rights reserved.

Keywords

  • Alzheimer Disease
  • Amyloid Precursor Protein Secretases
  • Base Sequence
  • Chromosome Mapping
  • Genetic Loci
  • Haplotypes
  • Humans
  • Membrane Glycoproteins
  • Molecular Sequence Data

Fingerprint Dive into the research topics of 'Functional and genetic analysis of haplotypic sequence variation at the nicastrin genomic locus'. Together they form a unique fingerprint.

Cite this