A protocol to study three-dimensional genome structure in individual mutant preimplantation mouse embryos

Summary: Hi-C studies the three-dimensional structure of the genome by detecting genome-wide chromatin regions that are in spatial proximity within the nucleus. We developed single-blastocyst Hi-C in mutant mouse embryos to genotype them on sequence. We describe steps for embryo fixation and nuclei...

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Bibliographic Details
Main Authors: Daniel Gimenez-Llorente, Marta Portela, Ana Losada, Miguel Manzanares, Maria Jose Andreu
Format: Article
Language:English
Published: Elsevier 2023-06-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166723002253
Description
Summary:Summary: Hi-C studies the three-dimensional structure of the genome by detecting genome-wide chromatin regions that are in spatial proximity within the nucleus. We developed single-blastocyst Hi-C in mutant mouse embryos to genotype them on sequence. We describe steps for embryo fixation and nuclei permeabilization, after which chromatin is digested and re-ligated having incorporated a biotin-labeled nucleotide at the ligation junction. After cross-link reversal, we then detail purification of immobilized chimeric DNA ligations, library generation, sequencing, and genome-wide analysis of interactions.For complete details on the use and execution of this protocol, please refer to Andreu et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667