Characterising open chromatin identifies novel cis-regulatory elements important for paraxial mesoderm formation and axis extension
| Autor: | Andrea Münsterberg, Victor Martinez-Heredia, Eirini Maniou, Ruth M. Williams, Gi Fay Mok, Grant N. Wheeler, Shannon A. Weldon, Leighton Folkes, Simon Moxon, Alice M. Godden |
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| Rok vydání: | 2020 |
| Předmět: |
Regulation of gene expression
0303 health sciences Cellular differentiation 030302 biochemistry & molecular biology Biology Paraxial mesoderm formation Chromatin Cell biology 03 medical and health sciences Somite medicine.anatomical_structure medicine Epigenome editing Paraxial mesoderm Hox gene 030304 developmental biology |
| DOI: | 10.1101/2020.01.20.912337 |
| Popis: | SUMMARYThe development of multicellular organisms is exquisitely regulated through differential gene activity, which governs cell differentiation programs. However, many details of spatiotemporal control of gene regulation are still poorly understood. We used the accessibility of chick embryos to examine genome-wide signatures characterizing the progressive differentiation of paraxial mesoderm along the head-to-tail axis. Paraxial mesoderm becomes organized into repetitive units, termed somites, the hallmark of the segmented vertebrate body plan. New somite pairs form periodically as the axis extends at the posterior end. This process generates a developmental gradient within a single embryo, with anterior somites more advanced in their differentiation compared to posterior somites. Following somite formation, cell rearrangements generate compartments, comprising lineages of the musculoskeletal system, including cartilage of the vertebral column and ribs, and skeletal muscle cells of the trunk and limbs. To examine how paraxial mesoderm becomes regionalized and patterned to eventually generate these discrete lineages, we investigated dynamic changes of the transcriptome and of chromatin accessibility using RNA-seq and ATAC-seq across a spatiotemporal series along the embryonic axis. Footprint analysis uncovers differential coverage of binding sites for a number of key transcription factors known to be involved in axial patterning and differentiation, including HOX genes. Furthermore, associating accessible chromatin with nearby expressed genes identifies candidatecis-regulatoryelements (CRE). As exemplars we use TCF15 and MEOX1, which are crucial for somite formation and differentiation, to experimentally validate CREsin vivousing fluorescent reporters. Time-lapse microscopy reveals CRE spatiotemporal activity and mutation analysis uncovers necessary upstream regulators. The CRE for MEOX1 is conserved and recognized in Xenopus. In addition, a human element is active in chicken.In vivoepigenome editing of TCF15 and MEOX1 CREs disrupts gene expression regulation and recapitulates phenotypic abnormalities of anterior-posterior axis extension. |
| Databáze: | OpenAIRE |
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