CAREER: Engineering principles within cell-cell communication networks in animal development

1254344细胞间信号的调节在生物学的许多领域都是至关重要的，包括发育、干细胞生物学和成年组织的适当维护。在发育过程中，组织是由长距离的、高度可变的信号组成的，这些信号指导着不同位置基因表达的差异。在首席研究员的实验室中，首要目标是将工程原理与调控机制联系起来，以确保在面临高度可变的细胞间信号时可靠的基因表达模式。这项拟议的工作旨在研究生物调节机制中工程原理的两个具体实例：反馈控制和噪声过滤。果蝇黑腹果蝇被用作模式生物，因为它在所需的复杂性(作为多细胞生物体)和易操作性之间取得了平衡，并配备了先进的遗传和转基因工具包。在早期(1-3小时大)的果蝇胚胎中，背侧-腹侧轴是由转录因子DRADAL形成的。在以前的工作中，人们发现背部即使在高内在噪声和外在变异性的情况下也建立了健壮的基因表达模式。与第二个信号通路，骨形态发生蛋白(BMP)通路的负反馈相互作用被认为是为了增加基因表达模式相对于背部信号变化的稳健性。此外，背部靶基因的表达位置远远超出了信号分子的空间梯度预期可靠发挥作用的范围。因此，将研究在噪声最高的背部信号尾部的基因表达的噪声过滤能力。负反馈相互作用和噪声过滤将使用遗传和转基因操作、基因表达输出的定量(实时)成像/模型、定量实时PCR和定量Western Blot进行研究。与拟议的实验研究相关，推广目标是通过强调生物系统中发现的工程原理的教育材料，促进工程学和生物学之间的协同作用。将与科学之家合作，为高中生开发和传播一个丰富的模块，科学之家是NCSU的一个组织，负责监督K-12的推广。目标是培养未来生物专业的高中生、初中生和高中生，熟悉工程学和物理学的语言和观点。由国际学生联合会教授的课程的本科生将作为助教参加。在研究生阶段，在NCSU远程教育计划的帮助下，将广泛传播一门专注于组织模式工程原理的专题课程。这些教育材料与研究工作一起，将为理解和利用自然工程提供视角和研究。存在于生物系统中。

1254344ReevesRegulation of signaling between cells is crucial in many areas of biology, including development, stem cell biology, and proper maintenance of adult tissues. In development, tissues are patterned by long range, yet highly variable signals that direct differences in gene expression at different locations. In the Principal Investigator's lab, the overarching goal is to connect engineering principles to the regulatory mechanisms that ensure reliable gene expression patterns in the face of highly variable intercellular signals. The proposed work aims to study two specific instances of engineering principles in biological regulatory mechanisms: feedback control and noise-filtering. The fruit fly Drosophila melanogaster is used as a model organism, as it strikes a balance between the complexity needed (as a multicellular organism) and ease of manipulation, with advanced genetic and transgenesis toolkits. In the early (1-3 hours old) Drosophila embryo, the dorsal-ventral axis is patterned by the transcription factor Dorsal. In previous work, it was found that Dorsal establishes robust gene expression patterns even in the face of high intrinsic noise and extrinsic variability. Negative feedback interactions with a secondary signaling pathway, the Bone Morphogenetic Protein (BMP) pathway, are hypothesized to increase the robustness of gene expression patterns with respect to variations in Dorsal signaling. Furthermore, dorsal target genes are expressed in a location far beyond the point in which a spatial gradient of a signaling molecule is expected to reliably act. Thus, the noise-filtering capabilities of gene expression in the tail of Dorsal signaling, where noise is the highest, will be investigated. Both negative feedback interactions and noise-filtering will be studied using genetic and transgenic manipulations, quantitative (live) imaging/models of gene expression outputs, quantitative real-time PCR and quantitative Western Blot.Related to the proposed experimental studies, the outreach goal is to facilitate the synergy between engineering and biology through educational materials that highlight the engineering principles found in biological systems. An enrichment module for high school students will be developed and disseminated in partnership with the Science House, an NCSU organization that oversees K-12 outreach. The goal is to train future biology majors, at the high school junior and senior level, to be comfortable with the language and viewpoint of engineering and physics. Undergraduate students in courses taught by the PI will participate as teaching assistants. At the graduate level, a special topics course that focuses on the engineering principles in tissue patterning will be broadly disseminated with the help of the NCSU Distance Education Program. These educational materials, together with the research work, will provide the perspective and research to understand and then utilize the ?natural engineering? present in biological systems.