Abstract
Cis-regulatory elements are critical for the precise spatiotemporal regulation of genes during development. However, identifying functional regulatory sites that drive cell differentiation in vivo has been complicated by the high numbers of cells required for whole-genome epigenetic assays. Here, we identified putative regulatory elements during sex determination by performing ATAC-seq and ChIP-seq for H3K27ac in purified XX and XY gonadal supporting cells before and after sex determination in mice. We show that XX and XY supporting cells initiate sex determination with similar chromatin landscapes and acquire sex-specific regulatory elements as they commit to the male or female fate. To validate our approach, we identified a functional gonad-specific enhancer downstream of Bmp2, an ovary-promoting gene. This work increases our understanding of the complex regulatory network underlying mammalian sex determination and provides a powerful resource for identifying non-coding regulatory elements that could harbor mutations that lead to Disorders of Sexual Development.
| Original language | English |
|---|---|
| Pages (from-to) | 168-179 |
| Number of pages | 12 |
| Journal | Developmental Biology |
| Volume | 446 |
| Issue number | 2 |
| DOIs | |
| State | Published - 15 Feb 2019 |
| Externally published | Yes |
Bibliographical note
Publisher Copyright:© 2019 The Authors
Funding
We thank the NU Sequencing core for their help with ChIP-seq and ATAC-seq, and the R. H. Lurie Comprehensive Cancer Center Flow Cytometry Facility for their help with FACS-sorting. We thank the Genetic Modification Service of the Francis Crick Institute for their help with the transient transgenics. We’d like to thank Peter van Galen for his help with the ChIP-seq protocol, and Lindsey Barske and Blanche Capel for their comments on the manuscript. Research was partially supported by the NIH to SAGM (NIH T32 GM008061 ), and IMS (NIH T32 DK007169 ) and partly by Francis Crick core funding ( Cancer Research UK , the U.K. Medical Research Council , and the Wellcome Trust : FC001107 ). We thank the NU Sequencing core for their help with ChIP-seq and ATAC-seq, and the R. H. Lurie Comprehensive Cancer Center Flow Cytometry Facility for their help with FACS-sorting. We thank the Genetic Modification Service of the Francis Crick Institute for their help with the transient transgenics. We'd like to thank Peter van Galen for his help with the ChIP-seq protocol, and Lindsey Barske and Blanche Capel for their comments on the manuscript. Research was partially supported by the NIH to SAGM (NIH T32 GM008061), and IMS (NIH T32 DK007169) and partly by Francis Crick core funding (Cancer Research UK, the U.K. Medical Research Council, and the Wellcome Trust: FC001107).
| Funders | Funder number |
|---|---|
| U.K. Medical Research Council | |
| National Institutes of Health | T32 GM008061 |
| National Institute of Diabetes and Digestive and Kidney Diseases | T32DK007169 |
| Wellcome Trust | FC001107 |
| Medical Research Council | |
| Cancer Research UK | |
| Institute for Molecular Science | T32 DK007169 |
Keywords
- Cell differentiation
- Chromatin landscapes
- Enhancers
- Gonad development
- Regulatory elements
- Sex determination