Dissecting complex genetic interaction networks mediating the fungal stress response

剖析介导真菌应激反应的复杂遗传相互作用网络

• 批准号: RGPIN-2018-04914
• 负责人: Shapiro, Rebecca
• 金额: $ 2.99万
• 依托单位: University of Guelph
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=664880
• 关键词: Dissecting; complex; genetic; interaction; networks

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.*****

微生物会遇到一系列的环境压力条件，包括养分可获得性、pH、温度和其他应激源的变化-它们的生存取决于它们感知、反应和适应这些变化的能力。一种这样的微生物是白色念珠菌：一种在健康的人类微生物群中常见的真菌，也是一种条件致病菌。白念珠菌会遇到并耐受一系列的应激条件，包括宿主内的温度波动和来自免疫系统的攻击。作为对压力的反应，白色念珠菌可以经历细胞形态发生程序，并从圆形的酵母形态转变为细长的丝状形态。为了了解白色念珠菌对压力的耐受和反应，我们必须确定控制其压力反应的遗传因素和遗传交互作用。我的研究计划将使用尖端基因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白念珠菌的应激反应？在不同的应激条件下，遗传交互作用是如何保守的？同样的遗传网络参与压力反应和形态发生吗？*我之前的工作建立了‘基因驱动阵列(GDA)’-一个基于CRISPR的平台，在白色念珠菌中创建单基因和双基因突变，并促进我们执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，使我们能够建立更大规模的白念珠菌基因突变文库。我们将使用这项技术来建立一个包含关键应激反应因子突变的白色念珠菌菌株文库--包括单个突变和双突变的每一种组合。该文库将在不同的压力条件下进行筛选，以确定控制生存和对压力反应的遗传因素和遗传交互作用。显微镜和成像分析将被用来确定压力条件和突变如何影响白色念珠菌的形态发生。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化技术来剖析细胞通路，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各种微生物中推广。