Biochemistry (Moscow)

, Volume 78, Issue 8, pp 895–903

Assay of insulator enhancer-blocking activity with the use of transient transfection

  • N. A. Smirnov
  • D. A. Didych
  • S. B. Akopov
  • L. G. Nikolaev
  • E. D. Sverdlov
Article

DOI: 10.1134/S0006297913080051

Cite this article as:
Smirnov, N.A., Didych, D.A., Akopov, S.B. et al. Biochemistry Moscow (2013) 78: 895. doi:10.1134/S0006297913080051

Abstract

We used a transient transfection of cultured cells with linearized plasmids to analyze the enhancer-blocking activity of potential insulators including the standard cHS4 chicken beta-globin insulator and several DNA fragments selected from the human genome sequence. About 60–80% of the potential insulators do reveal the enhancer-blocking activity when probed by the transient transfection assay. The activity of different sequences is characterized by certain tissue specificity and by dependence on the orientation of the fragments relative to the promoter. Thus, the transfection model may be used for quantitative analysis of the enhancer-blocking activity of the potential insulators.

Key words

human genome gene expression regulation insulator enhancer 

Copyright information

© Pleiades Publishing, Ltd. 2013

Authors and Affiliations

  • N. A. Smirnov
    • 1
  • D. A. Didych
    • 1
  • S. B. Akopov
    • 1
  • L. G. Nikolaev
    • 1
  • E. D. Sverdlov
    • 1
  1. 1.Shemyakin-Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia

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