Cell-free genomics: transcription factor interactions in reconstituted naïve embryonic chromatin

Extracts from Drosophila preblastoderm embryos (DREX) form the basis of a powerful in vitro chromatin reconstitution system that assembles entire genomes into complex chromatin with physiological nucleosome spacing and polymer condensation. As the zygotic genome has not yet been activated in preblas...

Full description

Saved in:
Bibliographic Details
Published inBiochemical Society transactions Vol. 52; no. 1; p. 423
Main Author Becker, Peter B
Format Journal Article
LanguageEnglish
Published England 28.02.2024
Subjects
Animals
Chromatin
Drosophila - metabolism
Genomics
Mammals - metabolism
Nucleosomes
Transcription Factors - metabolism
Drosophila preblastoderm embryo
nucleosome binding
pioneer transcription factor
nucleosome remodelling
chromatin immunoprecipitation
Online AccessGet more information

Cover

More Information
Summary:Extracts from Drosophila preblastoderm embryos (DREX) form the basis of a powerful in vitro chromatin reconstitution system that assembles entire genomes into complex chromatin with physiological nucleosome spacing and polymer condensation. As the zygotic genome has not yet been activated in preblastoderm embryos, the reconstitution extract lacks endogenous transcription factors (TFs) and the RNA polymerase machinery. At the same time, it contains high levels of ATP-dependent nucleosome sliding enzymes that render the reconstituted chromatin dynamic. The naïve chromatin can be used to determine the intrinsic DNA binding properties of exogenous, usually recombinant TFs (or DNA binding proteins in general) in a complex chromatin context. Recent applications of the system include the description of cooperation and competition of Drosophila pioneer TFs for composite binding sites, and the characterization of nucleosome interactions of mammalian pioneer TFs in the heterologous system.
ISSN:1470-8752
DOI:10.1042/BST20230878