Development of Bioengineered Cyanobacteriochromes for in vivo Fluorescence Tagging

蓝藻色素是一个蓝藻家族的胆囊蛋白光感受器。这一生物工程提案承诺一个小的、鲜红色的报告基因（约160个氨基酸）。与其他红色荧光标记物相比，胆道蛋白报告蛋白将表现出显著提高的光漂白抗性和低细胞内毒性。他们计划优化突变蓝藻色素的DNA序列，用于哺乳动物的表达，然后将其融合到哺乳动物的细胞骨架蛋白中，转染到活细胞中。提出的研究的智力优点是工程亮红色荧光蛋白是体内细胞成像的工具。目标是在感染期间成像红色荧光标记的细胞骨架，以便了解感染过程将提出治疗方案。明亮的红色，小的，无毒的荧光标签还有许多潜在的其他用途。该活动产生的更广泛的影响之一是培养生物工程方面的女本科生。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1名男性参加了米尔斯学院的生物光子学研究，该研究得到了美国国家科学基金会（CBST- UC Davis）的资助。这些学生都进入了研究生院或医学院。

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.