Pyrosequencing for accurate imprinted allele expression analysis

Bing Yang, Nathan Damaschke, Tianyu Yao, Johnathon McCormick, Jennifer Wagner, David Jarrard

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Genomic imprinting is an epigenetic mechanism that restricts gene expression to one inherited allele. Improper maintenance of imprinting has been implicated in a number of human diseases and developmental syndromes. Assays are needed that can quantify the contribution of each paternal allele to a gene expression profile. We have developed a rapid, sensitive quantitative assay for the measurement of individual allelic ratios termed Pyrosequencing for Imprinted Expression (PIE). Advantages of PIE over other approaches include shorter experimental time, decreased labor, avoiding the need for restriction endonuclease enzymes at polymorphic sites, and prevent heteroduplex formation which is problematic in quantitative PCR-based methods. We demonstrate the improved sensitivity of PIE including the ability to detect differences in allelic expression down to 1%. The assay is capable of measuring genomic heterozygosity as well as imprinting in a single run. PIE is applied to determine the status of Insulin-like Growth Factor-2 (IGF2) imprinting in human and mouse tissues. J. Cell. Biochem. 116: 1165-1170, 2015.

Original languageEnglish (US)
Pages (from-to)1165-1170
Number of pages6
JournalJournal of Cellular Biochemistry
Volume116
Issue number7
DOIs
StatePublished - Jul 1 2015
Externally publishedYes

Keywords

  • EPIGENETIC
  • IGF2
  • IMPRINTING
  • INTRON-CROSSING PRIMER
  • PYROSEQUENCING

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Yang, B., Damaschke, N., Yao, T., McCormick, J., Wagner, J., & Jarrard, D. (2015). Pyrosequencing for accurate imprinted allele expression analysis. Journal of Cellular Biochemistry, 116(7), 1165-1170. https://doi.org/10.1002/jcb.25081