title: Genetic investigation of FOXO3A requires special attention due to sequence homology with FOXO3B.
abstract: Our study demonstrates that the genetic investigation of forkhead box O3A gene (FOXO3A), a validated human longevity gene, is greatly hampered by the fact that its exonic regions have 99% sequence homology with the FOXO3B pseudogene. If unaccounted for, this high degree of homology can cause serious genotyping or sequencing errors. Here, we present an experimental set-up that allows reliable data generation for the highly homologous regions and that can be used for the evaluation of assay specificity. Using this design, we exemplarily showed FOXO3A-specific results for two single-nucleotide polymorphisms (SNPs) (rs4945816 and rs4946936) that are significantly associated with longevity in our centenarian-control sample (P(each)=0.0008). Because both SNPs are located in the 3' untranslated region of FOXO3A, they could be of functional relevance for the longevity phenotype. Our experimental set-up can be used for reliable and reproducible data generation for further sequencing and genotyping studies of FOXO3A with the aim of discovering new SNPs of functional relevance.
authors.affiliation: Institute of Clinical Molecular Biology, Christian-Albrechts-University, Schittenhelmstrasse 12,Kiel, Germany.
authors.forename: Friederike
authors.initials: F
authors.lastname: Flachsbart
authors.forename: Michael
authors.initials: M
authors.lastname: Möller
authors.forename: Carolin
authors.initials: C
authors.lastname: Däumer
authors.forename: Liljana
authors.initials: L
authors.lastname: Gentschew
authors.forename: Rabea
authors.initials: R
authors.lastname: Kleindorp
authors.forename: Michael
authors.initials: M
authors.lastname: Krawczak
authors.forename: Amke
authors.initials: A
authors.lastname: Caliebe
authors.forename: Stefan
authors.initials: S
authors.lastname: Schreiber
authors.forename: Almut
authors.initials: A
authors.lastname: Nebel
mesh_terms: D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008136:Longevity / Q000235:genetics*; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D011544:Pseudogenes / Q000235:genetics*; D017385:Sequence Homology
chemical_list: C499517:FOXO3 protein, human; D000071316:Forkhead Box Protein O3; D051858:Forkhead Transcription Factors