Discovery of craniofacial genes capable of compensation through evolutionary mutant model

脊椎动物的头面部骨骼是在高度的 脑神经脊细胞中活跃的保守基因调控网络。对这些进行了更改 网络可导致许多人类疾病，如腭裂、过早闭合 头骨，牙齿变小。对传统动物模型的实验室研究做出了贡献 对于我们理解核心基因网络的功能作用，但在很大程度上涉及 通过诱导突变进行显著的基因修改。因此，这些系统可能会更少 发现颅面基因网络如何适应基因丢失和独一无二 因为基因功能的丧失可能会产生显著的系统效应。一个 互补方法是研究具有适应性的进化突变模型 概述具有相似形态和/或基因变化的人类疾病。变异 对人类有害的可能是中性的，甚至在进化突变模型中是有益的， 因此，允许在正常生物体的背景下研究基因调控网络 功能。以前，有限的遗传工具需要在SELECT中检查头面部模型 动物模型。测序(如单细胞)和功能(如CRISPR)的最新进展 科技使对非传统物种的研究取得了丰硕成果。通过整合单细胞分析， 全基因组比较，以及功能分析，基因调控网络可以 成功地进行了跨物种的比较。我的项目将评估头面部基因网络 通过跨物种比较和研究合并体的保存性和延展性 鱼类(海豚、海马鱼和海豚)。这些令人惊叹的鱼已经拉长了筛骨 骨头，舌骨改变，牙齿完全脱落。另外，我们最近发现， 合并症丢失了我们推测的关键的头面部发育基因(fgf3和fgf4)。 导致了头面部基因调控网络的重新布线。首先，我将完成单个细胞 斑马鱼和刺鱼细胞RNA和染色质可及性的测序 (具有“正常”头面部特征的鱼类模型)和海豚(进化突变模型)。 其次，我将建立这些鱼和其他13种脊椎动物的全基因组比对，以确定 监管要素。第三，我将使用以下工具从功能上测试五个已确定的监管要素 斑马鱼。这三种方法将揭示颅面基因表达的保守性 模式和序列存在于众多物种中。此外，基因和序列 可能在适应基因丢失和产生改变的面部方面发挥作用，以及 可能会发现新的基因和调控因素，可以导致人类的治疗。

The craniofacial skeleton of vertebrates develops under the guidance of highly conserved gene regulatory networks active in cranial neural crest cells. Alterations to these networks can lead to numerous human disorders such as cleft palate, premature closing of the skull, and reduced teeth size. Laboratory studies of traditional animal models have contributed to our understanding of the functional role of the core gene networks but have largely involved significant gene modifications through induced mutations. As such, these systems may be less fruitful for discovering how craniofacial gene networks adapt to gene loss and unique morphologies because the abrogation of gene function can have a significant systemic effect. A complementary approach is the study of evolutionary mutant models with adaptations that recapitulate human diseases with similar altered morphology and/or gene changes. Variation that is detrimental in humans may be neutral or even beneficial in evolutionary mutant models, therefore allowing the study of gene regulatory networks in the context of normal organismal function. Previously, limited genetic tools necessitated examining craniofacial models in select animal models. Recent advances in sequencing (e.g. single cell) and functional (e.g. CRISPR) technologies enable fruitful studies in less traditional species. By integrating single cell analysis, whole genome comparisons, and functional assays, gene regulatory networks can be successfully compared across species. My project will evaluate craniofacial gene network conservation and malleability through cross species comparisons and studies of syngnathid fishes (pipefish, seahorses, and seadragons). These amazing fish have elongated ethmoid bones, altered hyoids, and a complete loss of teeth. In addition, we recently discovered that syngnathids have lost key craniofacial developmental genes (fgf3 and fgf4) that we hypothesize has led to rewiring of craniofacial gene regulatory networks. First, I will complete single cell sequencing to capture the RNA and chromatin accessibility of cells in zebrafish and stickleback (fish models with ‘normal’ craniofacial features), and pipefish (evolutionary mutant model). Second, I will build whole genome alignments of these fish and 13 other vertebrates to identify regulatory elements. Third, I will functionally test five identified regulatory elements using zebrafish. These three approaches will reveal how well conserved craniofacial gene expression patterns and sequences are across numerous species. In addition, genes and sequences unique to syngnathids may play a role in adaptation to gene loss and produce altered faces, and may identify novel genes and regulatory factors that can lead to human therapies.