Identifying the gene regulatory network of neurogenin - instrument supplement

项目摘要 精神分裂症是一种高度流行和衰弱的神经系统疾病，影响0.3%至0.6%的 美国人口。精神分裂症有很强的遗传成分，全基因组关联 研究已经确定了100多个导致这种疾病的候选基因座。在这份名单中， 因子神经生成素1。作为能够指导神经发生的前神经转录因子，该基因具有 在神经系统发育和功能过程中控制多个基因表达的潜力。尽管 这一点，很少有人知道的身份，neurogenin的下游转录目标，创造了一个必要的， 以供进一步研究。我们建议使用基因组学的方法来研究神经原蛋白周围的基因调控网络。 线虫C. elegans作为这些研究的一个易处理的模型。我们最近做了一个比较转录组 对C.线虫野生型和ngn-1/neurogenin突变体在胚胎发生中的作用。这表明，近600 在直接或间接的神经原转录控制下的基因。我们假设人类直系同源基因 的神经原转录靶点有很大的可能性是精神分裂症基因座在自己的权利。这 拟议的研究将使用染色质免疫沉淀加上下一代测序（ChIPseq）来识别 蠕虫基因组中的神经原蛋白结合位点，然后使用这些数据与我们的比较 转录组数据集，以描绘直接与间接的神经原转录靶点。本文书 补充者将购买浴式超声仪，以促进这些ChIPseq测定的染色质制备。我们将 使用GFP报告基因研究验证这些结果，重点是将神经生成素的控制置于环境中， 下游转录调节因子。最后，我们将使用最近发表的单细胞RNA测序， 数据集，再加上一个未发表的ngn-1表达谱系，以预测和验证基因，可能控制 ngn-1的转录。这种创新的方法将使上游调节剂作为神经元的一部分 基因调控网络这些基因的Homan直系同源物也可能有助于 精神分裂症该项目直接涉及神经系统发育的基本机制， 基因调控此外，它将实现广泛和具体的区域计划目标，包括提高 肯尼索州立大学的研究环境，并通过直接接触学生高质量的研究 参与的 PHS 398/2590（Rev. 11/07）第1页

Project Summary Schizophrenia is a highly prevalent and debilitating neurological disorder that affects between 0.3% and 0.6% of the U.S. population. There is a strong genetic component to schizophrenia, and Genome-Wide Association Studies have identified over 100 candidate loci that contribute to this disease. Among this list is the transcription factor neurogenin1. As a proneural transcription factor capable of directing neurogenesis, this gene has the potential to control the expression of multiple genes during nervous system development and function. Despite this, little is known about the identity of neurogenin’s downstream transcriptional targets, creating an imperative for further study. We propose to investigate the gene regulatory network surrounding neurogenin using the nematode C. elegans as a tractable model for these studies. We recently performed a comparative transcriptome study on C. elegans wild type and ngn-1/neurogenin mutants during embryogenesis. This identified nearly 600 genes that are under direct or indirect neurogenin transcriptional control. We hypothesize that human orthologs of neurogenin transcriptional targets have a strong possibility of being schizophrenia loci in their own right. This proposed study will use chromatin immunoprecipitation plus next-generation sequencing (ChIPseq) to identify neurogenin binding sites in the worm genome, then use these data to cross-reference against our comparative transcriptome dataset to delineate direct versus indirect neurogenin transcriptional targets. This instrument supplement will purchase a bath sonicator to facilitate chromatin preparation for these ChIPseq assays. We will validate these results using GFP reporter gene studies, with a focus on contextualizing neurogenin’s control of downstream transcriptional regulators. Finally, we will use recently published single cell RNA sequencing datasets, coupled with an unpublished ngn-1 expression lineage, to predict and validate genes that may control ngn-1’s transcription. This innovative approach will contextualize up-stream regulators as part of the neurogenin gene regulatory network. Homan orthologs of these genes may also contribute to the development of schizophrenia. This project directly addresses fundamental mechanisms of nervous system development and gene regulation. In addition, it will accomplish both broad and specific AREA program goals, including enhancing Kennesaw State University’s research environment and exposing students to high quality research through direct participation. PHS 398/2590 (Rev. 11/07) Page 1 Continuation Format Page