Technology for large-scale genetic interaction discovery in S. cerevisiae

描述（由申请人提供）：当两个基因同时受到干扰时，通常会出现令人惊讶的表型。遗传相互作用--由这种现象定义--表明相互作用的基因具有相关的功能。在这里，我们建议开发一种新的技术（“条形码融合遗传学”或BFG）检测遗传相互作用的S。酿酒。BFG方法利用携带条形码基因缺失的菌株的现有文库，并利用下一代测序技术的通量和经济性。如果成功的话，BFG技术有可能允许一名技术人员在一年内生成所有1800万S之间的遗传相互作用的地图。酿酒酵母基因对在任何给定的生长条件。在这项为期两年的试点计划中，我们建议开发和优化BFG技术，并通过将其应用于DNA修复和RNA聚合酶II转录延伸过程来评估其灵敏度和潜在价值。 如果两个基因的扰动一起产生令人惊讶的表型，则定义两个基因具有遗传相互作用。复杂的人类疾病，如癌症或糖尿病，需要多种突变，因此是遗传相互作用的结果。在这里，我们提出了一种技术，在模式生物S。酿酒酵母，可以经济地映射在给定的生长环境中的所有基因之间的遗传相互作用，并将该方法应用于DNA修复和转录基因的试点研究。

DESCRIPTION (provided by applicant): When two genes are perturbed simultaneously, a surprising phenotype often emerges. Genetic interaction - defined by this phenomenon - suggests that the interacting genes have related functions. Here we propose to develop a new technology ("bar-code fusion genetics" or BFG) for detecting genetic interactions in S. cerevisiae. The BFG method exploits existing libraries of strains carrying bar-coded gene deletions, and harnesses the throughput and economy of next-generation sequencing technology. If successful, the BFG technology has the potential to allow a single technician in a single year to generate a map of genetic interactions amongst all 18 million S. cerevisiae gene pairs in any given growth condition. In the context of this two-year pilot proposal, we propose to develop and optimize the BFG technology, and assess its sensitivity and potential value by applying it to the processes of DNA repair and RNA polymerase II transcription elongation. Two genes are defined to have a genetic interaction if the perturbation of both genes together yields a surprising phenotype. Complex human diseases such as cancer or diabetes require multiple mutations and are therefore the results of genetic interaction. Here we propose a technology in the model organism S. cerevisiae that could economically map genetic interactions amongst all genes in a given growth environment, and apply the approach in a pilot study of DNA repair and transcription genes.