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。由这一现象定义的基因相互作用表明相互作用的基因具有相关的功能。在此，我们建议开发一种新的技术（条形码融合遗传学）来检测酿酒葡萄球菌的遗传相互作用。BFG方法利用现有的携带条形码基因缺失的菌株文库，并利用下一代测序技术的吞吐量和经济性。如果成功的话，BFG技术有可能让一名技术人员在一年内绘制出在任何给定生长条件下所有1800万酿酒葡萄球菌基因对的基因相互作用图谱。在这个为期两年的试点项目中，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。

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.