Functional evolution of segmentation gene regulatory networks in insects

项目摘要 Evo-Devo领域的一个中心问题是控制胚胎发育的基因是如何在 进化论。Evo-Devo的许多最新进展已经确定了与Evo-Devo相关的基因变化 身体外部特征的获得或改变，如色素沉着模式或发育的改变 防弹衣。相比之下，我们的研究在这一领域是新颖的，因为它们揭示了意想不到的基因变异 其潜在的高度保守的特征是：昆虫共有的节段性身体结构。控制基因 片段编码胚胎发育和存活所需的转录因子。这群人 负责片段形成的基因包括在模式昆虫中发现的配对规则基因(PRGS) 果蝇。果蝇PRGS的突变会导致死亡，并伴随着交替节段区的丢失。 因此，令人惊讶的是，在不同的昆虫中发现了PRGS的存在、表达或功能的差异 分类群。这里提出的工作旨在了解这种遗传变异的机制基础，即 我们在不同的昆虫谱系中观察到。为了进行功能研究，我们开发了分子 在我们实验室的不同昆虫物种中的遗传方法。多个非模型系统的建立 由于共享协议和故障排除策略，在一个实验室内同时具有协同效应， 使我们能够在不同的物种中更有效地开发新技术。有了这些工具，我们将 研究特定遗传变异情景的潜在基础：在目标1中，我们将询问公关模式是如何 是在没有规范的PRGS的情况下实现的，我们在OnCopeltus中发现了 PRGS与果蝇的比较。目标2将探索OnCopeltus PRG网络的进化轨迹 跨越昆虫群。目标3将破译两种蚊子失去必要的PRG的潜在机制 还有蝴蝶。这些研究将有助于我们理解基本的调控机制 胚胎发育以及这些机制在昆虫辐射过程中是如何变化的。这个项目 将培养分子生物学的博士后研究员，研究生和至少五名本科生， 遗传学和分子进化。分子技术在非模式和新兴模式中的建立 昆虫种类，包括表达分析、RNA干扰、CRISPR/Cas9、FIRE-SEQ和转基因， 不仅允许我们回答关于胚胎发育的基本问题，而且还提供了分子 对对人类健康构成威胁的昆虫进行翻译研究的工具。

Project Summary A central question in the field of Evo-Devo is how genes controlling embryonic development change during evolution. Many recent advances in Evo-Devo have identified genetic changes that are associated with the acquisition of, or changes in, external body features, such as alterations in pigmentation patterns or development of body armor. In contrast, our studies are novel in this field as they have revealed unexpected genetic variation underlying a highly conserved trait: the shared segmented body plan of insects. The genes controlling segmentation encode transcription factors that are required for embryonic development and viability. The cohort of genes responsible for segment formation include pair-rule genes (PRGs) identified in the model insect Drosophila. Mutations in Drosophila PRGs result in lethality accompanied by loss of alternate segmental regions. Thus, it was surprising to find differences in the presence, expression, or function of PRGs in different insect taxa. The work proposed here is designed to understand the mechanistic basis for this genetic variation, which we have observed in different insect lineages. To carry out functional studies, we have developed molecular genetic approaches in diverse insect species in our lab. The establishment of multiple non-model systems simultaneously within one lab has synergistic effects due to sharing of protocols and troubleshooting strategies, allowing us to develop new techniques more effectively in different species. With these tools in hand, we will examine the underlying bases of specific scenarios of genetic variation: In Aim 1, we will ask how PR-patterning is achieved without canonical PRGs in the milkweed bug Oncopeltus where we have found novel utilization of PRGs compared to Drosophila. Aim 2 will explore the evolutionary trajectory of the Oncopeltus PRG network across insect groups. Aim 3 will decipher mechanisms underlying loss of an essential PRG in both mosquitoes and butterflies. These studies will contribute to our understanding of fundamental mechanisms regulating embryonic development and how these mechanisms have changed during the radiation of insects. This project will train postdoctoral fellows, graduate students, and at least five undergraduate students in molecular biology, genetics, and molecular evolution. Establishment of molecular techniques in non-model and emerging model insect species, including expression analysis, RNA interference, CRISPR/Cas9, FAIRE-seq, and transgenesis, not only allows us to answer fundamental questions about embryonic development, but also provides molecular tools for translational studies of insects that pose a risk to human health.