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

ABSTRACT: 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.

ORGANISM(S): Mus musculus

PROVIDER: GSE217197 | GEO | 2023/02/17

REPOSITORIES: GEO

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Project description:The Dlx homeobox genes have central roles in controlling patterning and differentiation of the brain and craniofacial primordia. In the brain, loss of Dlx function results in defects in the production, migration and differentiation of GABAergic neurons, that can lead to epilepsy. In the branchial arches, loss of Dlx function leads to craniofacial malformations that include trigeminal axon pathfinding defects. To determine how these genes function, we wish to identify the transcriptional circuitry that lies downstream of these transcription factors by comparing gene expression in wild type with Dlx mutant CNS and craniofacial tissues. 1) Compare gene expression in the maxillay branch of the first branchial arch (BA) of E10.5 wild type and Dlx2 -/- mutants. 2) Compare gene expression in the maxillary branch of the first BA of E10.5 wild type and Dlx1/2 -/- mutants. 3) Compare gene expression in wild type maxillary and mandibular branchial arches. 4) Compare gene expressionin mandibular branch of Dlx5/6 -/- mutants with wild type mandibular branch. The Dlx transcription factors are essential for controlling patterning of the brain and craniofacial primordia. In the brain, they control differentiation of GABAergic neurons of the basal ganglia. In the branchial arches, they control regional patterning. I hypothesize that there will be some conserved and some divergent mechanisms that the Dlx genes use in controlling brain and craniofacial development. We have already performed array analyses on Dlx function in the developing basal ganglia (with TGEN) by comparing expressed genes in wild type and Dlx1/2 mutants. Here we will compare gene expression in the brachial arches of wild type and Dlx mutant mice. 1) Generate E10.5 mouse embryos that are either wild type, Dlx2-/-, Dlx1/2 -/- or Dlx5/6 -/-. 2) Determine genotype by PCR. 3) Dissect branchial arches from the different genotypes. 4) Separate maxillary and mandibular branch of each branchial arch. 5) Prepare total RNA from the specimens. Obtain sufficient tissue to obtain 10 ug of total RNA - based on previous experience we anticipate that this will require ~ 10 branchial arches. We will pool the tissue from different embryos of the same genotype. 6) Send total RNA to TGEN for probe preparation, hybridization and array result analysis.

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

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