A Three-Dimensional In Vivo System for Visualizing Gene Expression

用于可视化基因表达的三维体内系统

• 批准号: 0236683
• 负责人: June Medford
• 金额: $ 10万
• 依托单位: Colorado State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-15 至 2007-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0236683&HistoricalAwards=false
• 关键词: Three; Dimensional; Vivo; System; Visualizing

How genes are expressed is a fundamental aspect of biology. An unmet challenge is to examine the three-dimensional gene expression patterns in intact organisms without disruption. Current technologies to follow gene expression in vivo either fail to penetrate deeply enough or provide too poor of a resolution for many basic research questions. Optical coherence microscopy (OCM) allows non-destructive, repetitive, quantitative, in vivo three-dimensional visualization of tissues and organs in intact organisms. Previous proof-of-concept experiments demonstrated that OCM could be used to non-destructively image in vivo, three-dimensional gene expression from a strong promoter using a naturally occurring polymer, PHB (poly-D-(-)-3-hydroxybutyrate) as the reporter gene. The goal of this project is to investigate the sensitivity of this method. Transgenic plants will be generated in which the gene reporter (PHB) is expressed at different levels and at different locations. These plants will then be analyzed using OCM to visualize the reporter gene product, and computer vision methods will be applied with the goal of determining the sensitivity of the method. Science, at its best, creates the future. A key part of how science "creates" the future is through discovery and understanding while educating others. Part of the support for the project will be for retention of a minority undergraduate in the Medford laboratory. In addition, partial support will be provided to two graduate students. To enhance the local impact of the work, an outreach activity to a local K-12 school is also planned. To enhance the wider educational impact of the work, information from the technology will be incorporated into classroom instruction. To enhance the wider scientific impact, presentations at professional meetings and publications in peer-reviewed journals are planned. This work will apply knowledge of engineers and computer vision specialists to develop the next-generation of gene analysis instruments for biologists. As such, this project is firmly positioned to significantly enhance scientific and technological understanding.

基因的表达方式是生物学的一个基本方面。 一个未满足的挑战是检查完整生物中的三维基因表达模式而不会破坏。 当前遵循体内基因表达的技术要么无法深入深度渗透，要么为许多基础研究问题提供了太差的分辨率。 光学相干显微镜（OCM）允许在完整生物体中对组织和器官的体内三维可视化无损，重复，定量。 先前的概念验证实验表明，使用天然存在的聚合物PHB（Poly-D - （ - （ - ） - 3-羟基丁酸酯）作为记者基因，使用了强启动子的体内，三维基因表达中OCM的三维基因表达。 该项目的目的是研究该方法的敏感性。 将产生转基因植物，其中基因报告基因（PHB）在不同的水平和不同位置表达。 然后，将使用OCM分析这些植物以可视化报告基因产物，并将应用计算机视觉方法以确定该方法的灵敏度。 科学最好地创造了未来。 科学如何创造未来的关键部分是通过发现和理解在教育他人的同时。 对该项目的部分支持将是保留梅德福实验室的少数本科生。 此外，将向两名研究生提供部分支持。 为了增强作品的当地影响，还计划向当地的K-12学校进行外展活动。 为了增强作品的更广泛的教育影响，该技术的信息将纳入课堂教学中。为了增强更广泛的科学影响，计划了在同行评审期刊上的专业会议和出版物的演讲。这项工作将应用工程师和计算机视觉专家的知识来开发生物学家基因分析工具的下一代。 因此，该项目牢固地定位，可以显着增强科学和技术理解。