Pyrosequencing for accurate imprinted allele expression analysis.

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.

journal_name

J Cell Biochem

authors

Yang B,Damaschke N,Yao T,McCormick J,Wagner J,Jarrard D

doi

10.1002/jcb.25081

subject

Has Abstract

pub_date

2015-07-01 00:00:00

pages

1165-70

issue

7

eissn

0730-2312

issn

1097-4644

journal_volume

116

pub_type

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