Abstract

Accumulating evidence suggests that the physical interactions between genomic regions play critical roles in the regulation of genome functions, such as transcription and epigenetic regulation. Various methods to detect the physical interactions between genomic regions have been developed. We recently developed a method to search for genomic regions interacting with a locus of interest in a non-biased manner that combines pull-down of the locus using engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) and next-generation sequencing (NGS) analysis (enChIP-Seq). The clustered regularly interspaced short palindromic repeats (CRISPR) system, consisting of a nuclease-dead form of Cas9 (dCas9) and a guide RNA (gRNA), or transcription activator-like (TAL) proteins, can be used for enChIP. In enChIP-Seq, it is necessary to compare multiple datasets of enChIP-Seq data to unambiguously detect specific interactions. However, it is not always easy to analyze enChIP-Seq datasets to subtract non-specific interactions or identify common interactions. To facilitate such analysis, we developed the enChIP-Seq analyzer software. It enables easy extraction of common signals as well as subtraction of non-specific signals observed in negative control samples, thereby streamlining extraction of specific enChIP-Seq signals. enChIP-Seq analyzer will help users analyze enChIP-Seq data and identify physical interactions between genomic regions.

Highlights

  • The dynamics of the three-dimensional structure of the genome play a critical role in the regulation of genome functions such as transcription, epigenetic regulation, genomic imprinting, and X-chromosome inactivation [1,2]

  • We recently developed a method to search for genomic regions interacting with a locus of interest in a non-biased manner that combines pull-down of the locus using engineered DNA-binding molecule-mediated chromatin immunoprecipitation and next-generation sequencing (NGS) analysis

  • It enables easy extraction of common signals as well as subtraction of non-specific signals observed in negative control samples, thereby streamlining extraction of specific engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP)-Seq signals. enChIP-Seq analyzer will help users analyze enChIP-Seq data and identify physical interactions between genomic regions

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Summary

Introduction

The dynamics of the three-dimensional structure of the genome play a critical role in the regulation of genome functions such as transcription, epigenetic regulation, genomic imprinting, and X-chromosome inactivation [1,2]. We recently developed a method to search for genomic regions interacting with a locus of interest in a non-biased manner that combines pull-down of the locus using engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) and next-generation sequencing (NGS) analysis (enChIP-Seq). It is not always easy to analyze enChIP-Seq datasets to subtract non-specific interactions or identify common interactions. To facilitate such analysis, we developed the enChIP-Seq analyzer software. It enables easy extraction of common signals as well as subtraction of non-specific signals observed in negative control samples, thereby streamlining extraction of specific enChIP-Seq signals. EnChIP-Seq analyzer will help users analyze enChIP-Seq data and identify physical interactions between genomic regions It enables easy extraction of common signals as well as subtraction of non-specific signals observed in negative control samples, thereby streamlining extraction of specific enChIP-Seq signals. enChIP-Seq analyzer will help users analyze enChIP-Seq data and identify physical interactions between genomic regions

Materials and Methods
Procedures for enChIP-Seq
Filtering of the NGS Peaks to Identify Genomic Regions Interacting with a Target
Preparation of the Data
HExantrdalcintigoonf othfecSoymstmemon peak information
Elimination of Negative Peak Information (2-1)
Extraction of Peak Information Commonly Included in Two or More Tab Files (1-1)
View Information within a Tab File (1)
Results and Discussion
Conclusions
Availability and Requirements
Full Text
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