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在发育过程中如何变化。 进化最近在Evo-Devo方面的许多进展已经确定了与发育相关的遗传变化。 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遗传变异 这是一个高度保守的特征：昆虫共有的分段身体结构。基因控制 分割编码胚胎发育和存活所需的转录因子。队列 包括在模式昆虫中鉴定的成对规则基因（PRG 果蝇果蝇PRG的突变导致致死性，伴随着交替节段区域的丢失。 因此，令人惊讶的是，在不同的昆虫中发现PRG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， 我们在不同的昆虫谱系中观察到的。为了进行功能研究，我们开发了分子 遗传学方法在我们实验室的不同昆虫物种中。多个非模型系统的建立 由于共享协议和故障排除策略， 使我们能够在不同物种中更有效地开发新技术。有了这些工具，我们将 检查遗传变异的特定场景的潜在基础：在目标1中，我们将询问PR模式如何 在乳草属昆虫Oncopeltus中没有典型的PRG，我们发现了新的利用 PRG与果蝇相比。目标2将探索Oncopeltus PRG网络的进化轨迹 在昆虫群体中。目标3将解释两种蚊子中基本PRG丢失的潜在机制 还有蝴蝶这些研究将有助于我们理解调节 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 将培养博士后研究员、研究生和至少5名分子生物学本科生， 遗传学和分子进化。非模型和新兴模型中分子技术的建立 昆虫物种，包括表达分析，RNA干扰，CRISPR/Cas9，FAIRE-seq和转基因， 不仅可以让我们回答有关胚胎发育的基本问题， 对人类健康构成风险的昆虫的转化研究工具。

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.