Pyrosequencing for accurate imprinted allele expression analysis.
Pyrosequencing for accurate imprinted allele expression analysis.
复制标题
焦磷酸测序用于准确的印记等位基因表达分析。
DOI：
10.1002/jcb.25081
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发表时间：
2015
影响因子：
4
通讯作者：
Jarrard,David
中科院分区：
文献类型：
--
作者：
Yang,Bing;Damaschke,Nathan;Yao,Tianyu;McCormick,Johnathon;Wagner,Jennifer;Jarrard,David
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 ofInsulin‐like Growth Factor‐2(IGF2) imprinting in human and mouse tissues. J. Cell. Biochem. 116: 1165–1170, 2015. © 2015 Wiley Periodicals, Inc.
