Методы высокопроизводительного репортерного анализа энхансеров

Translated title of the contribution: Methods of massive parallel reporter assays for investigation of enhancers

Research output: Contribution to journalArticlepeer-review

Abstract

The correct deployment of genetic programs for development and differentiation relies on finely coordinated regulation of specific gene sets. Genomic regulatory elements play an exceptional role in this process. There are few types of gene regulatory elements, including promoters, enhancers, insulators and silencers. Alterations of gene regulatory elements may cause various pathologies, including cancer, congenital disorders and autoimmune diseases. The development of high-throughput genomic assays has made it possible to significantly accelerate the accumulation of information about the characteristic epigenetic properties of regulatory elements. In combination with high-throughput studies focused on the genome-wide distribution of epigenetic marks, regulatory proteins and the spatial structure of chromatin, this significantly expands the understanding of the principles of epigenetic regulation of genes and allows potential regulatory elements to be searched for in silico. However, common experimental approaches used to study the local characteristics of chromatin have a number of technical limitations that may reduce the reliability of computational identification of genomic regulatory sequences. Taking into account the variability of the functions of epigenetic determinants and complex multicomponent regulation of genomic elements activity, their functional verification is often required. A plethora of methods have been developed to study the functional role of regulatory elements on the genome scale. Common experimental approaches for in silico identification of regulatory elements and their inherent technical limitations will be described. The present review is focused on original high-throughput methods of enhancer activity reporter analysis that are currently used to validate predicted regulatory elements and to perform de novo searches. The methods described allow assessing the functional role of the nucleotide sequence of a regulatory element, to determine its exact boundaries and to assess the influence of the local state of chromatin on the activity of enhancers and gene expression. These approaches have contributed substantially to the understanding of the fundamental principles of gene regulation.

Translated title of the contributionMethods of massive parallel reporter assays for investigation of enhancers
Original languageRussian
Article number12
Pages (from-to)344-355
Number of pages12
JournalVavilovskii Zhurnal Genetiki i Selektsii
Volume25
Issue number3
DOIs
Publication statusPublished - 2021

Keywords

  • Enhancers
  • Gene regulatory elements
  • Massive parallel assays

OECD FOS+WOS

  • 3 MEDICAL AND HEALTH SCIENCES
  • 1.06 BIOLOGICAL SCIENCES
  • 1.06.KM GENETICS & HEREDITY

State classification of scientific and technological information

  • 34 BIOLOGY

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