CAREER: A Cell-Based Biosensor for DNA Damaging Agents

这项研究的目的是建立基于细胞的DNA损伤剂传感器。真核细胞已经进化出非常敏感和快速的机制来感知和响应DNA损伤。该项目的第一阶段将定量确定芽殖酵母酿酒酵母中DNA损伤敏感基因转录对一组选定的损伤剂的剂量和时间依赖性。对DNA损伤剂敏感和选择性的基因簇将作为生物传感器设计的基础。接下来，损伤调节基因将被报告基因取代，以便于测量基因转录。该项目的第三阶段将通过禁用内源性DNA损伤途径来增强细胞对DNA损伤的敏感性。最后，该项目将为工程本科生和高中生创造分子和细胞生物工程领域的新实验室材料。这些实验室将展示通过改变细胞的遗传组成来定量控制细胞如何对环境刺激做出反应的能力。

The objective of this proposed research is the creation of cell-based sensors of DNA-damaging agents. Eukaryotic cells have evolved extremely sensitive and rapid mechanisms to sense and respond to DNA damage. The first stage of this project will quantitatively determine the dose and time-dependence of DNA damage-sensitive gene transcription in the budding yeast Saccharomyces cerevisiae in response to a panel of selected damaging agents. Clusters of genes sensitive and selective for DNA-damaging agents will serve as the basis for biosensor design. Next, the damage-regulated genes will be replaced with reporter genes to facilitate measurement of gene transcription. The third stage of the project will enhance the sensitivity of the cells to DNA damage by disabling endogenous DNA damage pathways. Finally, this project will create new laboratory materials in molecular and cellular bioengineering for undergraduate engineering and high school students. The labs will illustrate the ability to quantitatively control how cells respond to environmental stimuli by altering the genetic makeup of the cells.