Maatouk Danielle M, Natarajan Anirudh, Shibata Yoichiro, Song Lingyun, Crawford Gregory E, Ohler Uwe, Capel Blanche
Department of Cell Biology, Duke University, Durham, NC 27710, USA.
Department of Obstetrics and Gynecology, Northwestern University, Chicago, IL 60611, USA.
Development. 2017 Feb 15;144(4):720-730. doi: 10.1242/dev.142554. Epub 2017 Jan 13.
A current goal of molecular biology is to identify transcriptional networks that regulate cell differentiation. However, identifying functional gene regulatory elements has been challenging in the context of developing tissues where material is limited and cell types are mixed. To identify regulatory sites during sex determination, we subjected Sertoli cells from mouse fetal testes to DNaseI-seq and ChIP-seq for H3K27ac. DNaseI-seq identified putative regulatory sites around genes enriched in Sertoli and pregranulosa cells; however, active enhancers marked by H3K27ac were enriched proximal to only Sertoli-enriched genes. Sequence analysis identified putative binding sites of known and novel transcription factors likely controlling Sertoli cell differentiation. As a validation of this approach, we identified a novel Sertoli cell enhancer upstream of , and used it to drive expression of a transgenic reporter in Sertoli cells. This work furthers our understanding of the complex genetic network that underlies sex determination and identifies regions that potentially harbor non-coding mutations underlying disorders of sexual development.
分子生物学当前的一个目标是识别调控细胞分化的转录网络。然而，在发育中的组织中，由于材料有限且细胞类型混合，识别功能性基因调控元件一直具有挑战性。为了在性别决定过程中识别调控位点，我们对来自小鼠胎儿睾丸的支持细胞进行了DNaseI测序和H3K27ac的ChIP测序。DNaseI测序在支持细胞和前颗粒细胞中富集的基因周围识别出了假定的调控位点；然而，以H3K27ac标记的活性增强子仅在支持细胞富集的基因近端富集。序列分析确定了可能控制支持细胞分化的已知和新型转录因子的假定结合位点。作为这种方法的验证，我们在[具体基因]上游鉴定了一个新型的支持细胞增强子，并用它来驱动转基因报告基因在支持细胞中的表达。这项工作进一步加深了我们对性别决定基础的复杂遗传网络的理解，并识别出可能存在性发育障碍潜在非编码突变的区域。
