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Molecular Biology

, Volume 53, Issue 3, pp 346–353 | Cite as

Relative Efficiency of Transcription Factor Binding to Allelic Variants of Regulatory Regions of Human Genes in Immunoprecipitation and Real-Time PCR

  • N. A. Mitkin
  • K.V. Korneev
  • A. M. Gorbacheva
  • D. V. KuprashEmail author
GENOMICS. TRANSCRIPTOMICS
  • 13 Downloads

Abstract

The efficiency at which specific transcription factors interact with DNA may vary in the presence of single nucleotide polymorphisms (SNPs), and the variation provides an important mechanism that regulates expression of human genes and contributes to the individual susceptibility to various diseases. Ample genetic and epigenetic data make it possible to predict both functional polymorphic variants and the transcription factors whose binding they affect. However, predictions of the kind require experimental verification. An original method developed for the purpose includes immunoprecipitation of DNA–protein complexes, followed by quantification of the bound DNA by real-time PCR. The method does not require chemical modification of the DNA probes and yield reproducible results with total nuclear extracts from cultured human cells.

Keywords:

immunoprecipitation DNA–protein interactions transcription factors 

Notes

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Copyright information

© Pleiades Publishing, Inc. 2019

Authors and Affiliations

  • N. A. Mitkin
    • 1
  • K.V. Korneev
    • 1
  • A. M. Gorbacheva
    • 1
    • 2
  • D. V. Kuprash
    • 1
    • 2
    Email author
  1. 1.Engelhardt Institute of Molecular Biology, Russian Academy of SciencesMoscowRussia
  2. 2.Biological Faculty, Moscow State UniversityMoscowRussia

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