Development of Bioengineered Cyanobacteriochromes for in vivo Fluorescence Tagging

用于体内荧光标记的生物工程蓝藻色素的开发

• 批准号: 0967965
• 负责人: Susan Spiller
• 金额: $ 38.61万
• 依托单位: Mills College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-15 至 2014-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0967965&HistoricalAwards=false
• 关键词: Development; Bioengineered; Cyanobacteriochromes; vivo; Fluorescence

0967965SpillerCyanobacteriochromes are a cyanobacterial family of biliprotein photoreceptors. This bioengineering proposal promises a small, bright red reporter (about 160 amino acids). Compared to other red-fluorescent markers, biliprotein reporters will exhibit substantially improved resistance to photobleaching and low intracellular toxicity. They plan to optimize the DNA sequence of a mutant cyanobacteriochrome for mammalian expression, and then to fuse it to mammalian cytoskeleton proteins for transfection into living cells. The intellectual merit of the proposed research is that engineered bright red fluorescent proteins are tools for in vivo cellular imaging. The goal is to image a red fluorescent-tagged cytoskeleton during infection so that understanding the infection process will suggest therapeutic protocols. There are potentially many other uses for a bright red, small, non-toxic fluorescent tag. Among the broader impacts resulting from the activity is the training of undergraduate women in bioengineering. Mills College's small college atmosphere is an ideal environment for engaging young women in the excitement of scientific research. Twenty-eight young women and one man have participated in biophotonics research at Mills College supported by a sub award from NSF (CBST- UC Davis). All of these students have entered graduate schools or medical schools.

0967965 SpillerCyanobacteriochromes是胆蛋白光感受器的蓝细菌家族。这个生物工程的建议承诺一个小的，鲜红色的记者（约160个氨基酸）。与其他红色荧光标记物相比，胆蛋白报告基因将表现出显著改善的耐光漂白性和低细胞内毒性。他们计划优化突变蓝藻色素的DNA序列，使其在哺乳动物中表达，然后将其融合到哺乳动物细胞骨架蛋白中，以便转染到活细胞中。这项研究的智力价值在于，工程化的亮红色荧光蛋白是体内细胞成像的工具。目标是在感染期间对红色荧光标记的细胞骨架进行成像，以便了解感染过程将提出治疗方案。亮红色、小型、无毒的荧光标签还有许多其他潜在用途。该活动产生的更广泛影响之一是对大学女生进行生物工程方面的培训。米尔斯学院的小型学院氛围是吸引年轻女性参与科学研究的理想环境。28名年轻女性和一名男性参加了米尔斯学院的生物光子学研究，该研究得到了NSF（CBST-加州大学戴维斯分校）的子奖项的支持。所有这些学生都进入了研究生院或医学院。