Dissecting complex genetic interaction networks mediating the fungal stress response

微生物会遇到一系列环境胁迫条件，包括营养物质可用性、pH值、温度和其他胁迫因素的变化，它们的生存取决于它们感知、响应和适应这些变化的能力。其中一种微生物是白色念珠菌：一种常见于健康人体微生物组的真菌，也是一种机会致病菌。C.白色念珠菌遇到并耐受一系列应激条件，包括宿主体内的温度波动和免疫系统的攻击。在应激反应中，C.白色念珠菌可经历细胞形态发生程序，并从圆形酵母形态转变为细长的丝状形式。了解C.白念珠菌耐受压力并对压力作出反应，我们必须确定控制其压力反应的遗传因素和遗传相互作用。我的研究计划将使用尖端的遗传技术来解决真菌生物学中的基本问题：哪些遗传因素和遗传相互作用调节C。白色念珠菌应激反应在不同的胁迫条件下，遗传相互作用是如何保持的？应激反应和形态发生是否也涉及相同的遗传网络？我之前的工作建立了基因驱动阵列（GDA），这是一个基于CRISPR的平台，可以在C中创建单基因和双基因突变体。白念珠菌，并促进我们进行复杂的遗传相互作用分析的能力。在此基础上，我提出的研究计划将修改GDA系统，使我们能够构建C。白念珠菌基因突变体文库的研究。我们将使用这种技术来构建一个C库。白色念珠菌菌株在关键应激反应因子中含有突变，包括单独突变和双重突变的每种组合。该文库将在不同的应激条件下进行筛选，以确定控制生存和应激反应的遗传因素和遗传相互作用。显微镜和成像分析将用于确定应力条件和突变如何影响C。白色念珠菌我们将使用计算分析来构建遗传相互作用网络，并使用分子遗传学和生物化学技术来剖析细胞途径，并确定这些因素控制应激反应的机制。这项工作对我们理解真菌应激反应具有重要意义，我们开发的技术将在多种微生物中推广。

Microbial organisms encounter a range of environmental stress conditions, including changes in nutrient availability, pH, temperature, and other stressors and their survival is contingent on their abilities to sense, respond, and adapt to these changes. One such micro-organism is Candida albicans: a fungus commonly found in the healthy human microbiome, and an opportunistic pathogen. C. albicans encounters and tolerates a range of stress conditions, including temperature fluctuations within hosts, and assaults from the immune system. In response to stress, C. albicans can undergo a cellular morphogenesis program, and transition from a round, yeast morphology, to an elongated filamentous form. To understand how C. albicans tolerates and responds to stress, we must identify the genetic factors and genetic interactions that govern their stress response. My research program will use cutting-edge genetic technologies to address fundamental questions in fungal biology: which genetic factors and genetic interactions regulate the C. albicans stress response? How are genetic interactions conserved across different stress conditions? Are the same genetic networks involved in stress response and morphogenesis? My previous work established the Gene Drive Array (GDA)' a CRISPR-based platform to create single and double genetic mutants in C. albicans and facilitate our ability to perform complex genetic interaction analysis. Building on this, my proposed research program will modify the GDA system to allow us to build C. albicans genetic mutant libraries on a larger scale. We will use this technology to build a library of C. albicans strains containing mutations in key stress response factors both singly, and in every combination of double mutations. The library will be screened under diverse stress conditions, to identify genetic factors and genetic interactions that govern survival and response to stress. Microscopic and imaging analysis will be used to determine how stress conditions and mutations influence morphogenesis in C. albicans. We will use computational analysis to build genetic interaction networks, and use molecular genetic and biochemical techniques to dissect cellular pathways, and determine the mechanisms by which these factors govern the stress response. This work has important implications for our understanding of the fungal stress response, and the technologies we develop will be generalizable across a diversity of microbial organisms.