Effects of Dlx2 Overexpression on the Genome of Maxillary Process in Early Embryo of Mice [scRNA-seq]

Transcription factor Dlx2 plays an important role in craniomaxillofacial development. Overexpression or null mutation can lead to craniomaxillofacial malformation in mice. Although some in vivo or in vitro experiments have explored part information about the mechanism of Dlx2 regulation, there is still a lack of complete description. Using a mouse model that can stably overexpress Dlx2 in neural crest cells, the authors conducted bulk RNA-Seq, scRNA-Seq and CUT&Tag on the early maxillary processes of mice, and comprehensively described the effects of Dlx2 overexpression on the early development of maxillary processes. Bulk RNA-Seq results showed that Dlx2 had greater interference on nerve development at E10.5, and gradually affected bone development at E12.5. ScRNA-Seq proved that overexpression of Dlx2 did not change the differentiation type of mesenchymal cells during this development process, but it would restrict the proliferation of cells and make mesenchymal cells differentiate prematurely, thus limiting the development of maxillary. CUT&Tag suggested that this regulatory process is closely related to Notch signaling pathway, and MNT and RUNX2, as direct downstream regulatory target genes of Dlx2, also play a very critical role. Using a Dlx2 overexpression mouse model, the authors conducted bulk RNA Seq on the maxillary process of E10.5 mice, analyzed the impact of Dlx2 overexpression on genome expression, and compared it with the existing data of E12.5 mouse maxillary process in GEO database. Single cell RNA sequencing (scRNA Seq) was performed on the maxillary process of E12.5 mice, and the effects of overexpression of Dlx2 on cell population, cell cycle and developmental trajectory were analyzed. Cleavage under targets and segmentation (CUT&Tag) was performed on the maxillary process of E12.5 mice, the possible binding sites of DLx2 were analyzed, and the downstream target genome was searched by combining with bulk RNA Seq data.