CAREER: A Cell-Based Biosensor for DNA Damaging Agents

事业：基于细胞的 DNA 损伤剂生物传感器

• 批准号: 0238680
• 负责人: Sean Palecek
• 金额: $ 40万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0238680&HistoricalAwards=false
• 关键词: CAREER; Cell; Based; Biosensor; DNA

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.

这项拟议的研究的目标是创造基于细胞的DNA损伤剂传感器。真核细胞已经进化出极其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地确定酿酒酵母萌芽中DNA损伤敏感基因转录的剂量和时间依赖关系，以响应一组选定的损伤剂。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。下一步，损伤调控基因将被报告基因取代，以便于基因转录的测量。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，该项目将为工科本科生和高中生创造分子和细胞生物工程方面的新实验材料。这些实验室将展示通过改变细胞的基因组成来定量控制细胞对环境刺激的反应的能力。