title: Modeling Genomic Imprinting Disorders Using Induced Pluripotent Stem Cells.
abstract: Induced pluripotent stem cell (iPSC) technology has allowed for the invaluable modeling of many genetic disorders including disorders associated with genomic imprinting. Genomic imprinting involves differential DNA and histone methylation and results in allele-specific gene expression. Most of the epigenetic marks in somatic cells are erased and reestablished during the process of reprogramming into iPSCs. Therefore, in generating models of disorders associated with genomic imprinting, it is important to verify that the imprinting status and allele-specific gene expression patterns of the parental somatic cells are maintained in their derivative iPSCs. Here, we describe three techniques: DNA methylation analysis, allele-specific PCR, and RNA FISH, which we use to analyze genomic imprinting in iPSC models of neurogenetic disorders involving copy number variations of the chromosome 15q11-q13 region.
authors.affiliation: University of Connecticut Health Center, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA. chamberlain@uchc.edu.
authors.forename: Stormy J
authors.initials: SJ
authors.lastname: Chamberlain
authors.affiliation: University of Connecticut Health Center, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA.
authors.forename: Noelle D
authors.initials: ND
authors.lastname: Germain
authors.affiliation: University of Connecticut Health Center, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA.
authors.forename: Pin-Fang
authors.initials: PF
authors.lastname: Chen
authors.affiliation: University of Connecticut Health Center, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA.
authors.forename: Jack S
authors.initials: JS
authors.lastname: Hsiao
authors.affiliation: University of Connecticut Health Center, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA.
authors.forename: Heather
authors.initials: H
authors.lastname: Glatt-Deeley
mesh_terms: D000483:Alleles; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D056915:DNA Copy Number Variations; D019175:DNA Methylation; D017931:DNA Primers / Q000138:chemical synthesis / Q000378:metabolism; D044127:Epigenesis, Genetic; D061252:Feeder Cells / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D018392:Genomic Imprinting; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008957:Models, Genetic; D016133:Polymerase Chain Reaction / Q000379:methods; D011218:Prader-Willi Syndrome / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D012313:RNA / Q000235:genetics / Q000378:metabolism; D020537:RNA, Small Nucleolar / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism
chemical_list: D017931:DNA Primers; D020537:RNA, Small Nucleolar; D012313:RNA; C471639:UBE3A protein, human; D044767:Ubiquitin-Protein Ligases
keywords: Allele-specific PCR; DNA methylation; Genomic imprinting; Induced pluripotent stem cells; RNA FISH