Systematic Genetic Analysis with Conditional Estradiol-Regulated Gene Expression Alleles in Yeast

酵母条件雌二醇调节基因表达等位基因的系统遗传分析

• 批准号: RGPIN-2016-05539
• 负责人: Boone, Charles
• 金额: $ 2.77万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2016
• 资助国家: 加拿大
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=615757
• 关键词: Systematic; Genetic; Analysis; Conditional; Estradiol

The sophisticated molecular genetic methods available with yeast, makes this organism an ideal model system for studying fundamental cellular functions. The proposed project involves the construction of a new genome-wide collection of yeast mutants, in which each one of its ~6000 different genes is placed under the control of an inducible promoter, which means we can turn the expression of the gene on or off, simply by including or removing an inducer from the growth medium. This new mutant collection, which we refer to as the Z3 collection, has numerous applications and thus it will fuel research in numerous different labs around the world. We are particularly interested in this new mutant collection because it will allow us to track how cells die when particular genes are turned off. Interestingly, only ~1000 yeast genes are essential, such that the cell will not grow or divide when they are turned off or deleted, the other ~5000 genes are nonessential and the cell remains alive even when they are deleted or shut off. We think most genes are nonessential because cells are buffered from genetic and environmental perturbations and thus they have evolved backup genes and pathways to compensate for defects in a single gene. In another project, our group has tested all possible yeast double mutants for a lethal phenotype. While we now know many gene pairs work together to control cellular life, at a mechanistic level, we do not know exactly why the cell depends on both genes. We will use the new Z3 collection to investigate the terminal phenotype of a select set of yeast essential genes and lethal double mutants. We have assembled a number of diagnostic markers that allow us to follow the function of pathways and cellular compartments. We anticipate that different terminal phenotype trajectories will provide us with new knowledge of how genes and their corresponding pathways are connected to one another and thus will provide us with valuable new insights on how cells work.

酵母的复杂分子遗传学方法使这种生物体成为研究基本细胞功能的理想模型系统。拟议的项目涉及构建一个新的全基因组酵母突变体集合，其中约6000个不同基因中的每一个都置于诱导型启动子的控制下，这意味着我们可以打开或关闭基因的表达，简单地通过包括或从生长培养基中去除诱导剂。这个新的突变体集合，我们称之为Z3集合，有许多应用，因此它将推动世界各地许多不同实验室的研究。我们对这个新的突变体集合特别感兴趣，因为它将使我们能够跟踪当特定基因关闭时细胞如何死亡。 有趣的是，只有约1000个酵母基因是必需的，当它们被关闭或删除时，细胞将不会生长或分裂，其他约5000个基因是非必需的，即使它们被删除或关闭，细胞仍然活着。我们认为大多数基因都是非必需的，因为细胞可以从遗传和环境干扰中得到缓冲，因此它们已经进化出备份基因和途径来弥补单个基因的缺陷。在另一个项目中，我们的小组已经测试了所有可能的酵母双突变体的致命表型。虽然我们现在知道许多基因对共同控制细胞的生命，但在机械水平上，我们并不确切知道为什么细胞依赖于这两个基因。我们将使用新的Z3收集研究一组选择的酵母必需基因和致死性双突变体的终端表型。我们已经组装了一些诊断标记物，使我们能够跟踪通路和细胞区室的功能。我们预计，不同的终末表型轨迹将为我们提供有关基因及其相应通路如何相互连接的新知识，从而为我们提供有关细胞如何工作的有价值的新见解。