Enhancing and expanding the CGC Strain Collection

项目摘要/摘要--应用研究部分 线虫遗传中心(CGC)致力于提高科学家制造 生物医学研究的进展使用了一种主要的模式生物--秀丽线虫。转基因技术分析 动物基本上是所有模式生物研究中不可或缺的一部分。荧光蛋白(FP)简介 诺贝尔奖获得者马丁·查尔菲于1994年使用线虫对动物模型进行了标记，这是一场深刻的革命 转基因通过允许实时监测活生物体中的基因表达来实现。对于低基因 水平表达，荧光蛋白融合的检测可能是具有挑战性的，推动人们对更明亮的兴趣 以及荧光蛋白的光稳定变异。此外，许多应用程序需要多路传输 荧光标记的蛋白质，增加了对发射光谱不重叠的新蛋白质的需求。因此， 第一个绿色荧光蛋白的开发启发了许多变体的发现和设计，这些变体 可以在颜色、亮度、光稳定性和成熟时间上有所不同。但是，这些特征可以是 受环境条件的影响，以至于体外特性在体内可能不成立，这对 研究人员希望扩大他们的FP调色板。当一个新的FP被开发出具有高度吹捧的特性时， 体外特征参数允许选择良好的候选者进行体内测试，但采用了一种新的 蠕虫群落中的蛋白质需要进行重大的优化和体内测试。这个项目将消除 通过直接定量研究线虫群落新融合蛋白在体内使用的不确定性 一组新的FP彼此之间的比较以及与目前广泛使用的FP的比较。由此派生的数据 该项目将被线虫社区广泛使用，以改进多样化研究的实验设计 问题，以及研究人员使用其他模式生物建议在体内进行测试的最佳候选FP 他们的系统。该项目还将产生一组表达FP的线虫菌株，供实验室测试 他们自己的显微镜和/或他们正在考虑购买的演示设备上。最后，这个项目 将建立一套用于评估未来FP的通用指标，以确保 测试。在这项工作中产生的所有菌株和试剂将由CGC进行管理，并立即提供给 社区。

Project Summary/Abstract—Applied Research Component The Caenorhabditis Genetics Center (CGC) seeks to enhance the efficiency with which scientists can make biomedical research advances using a premier model organism, Caenorhabditis elegans. Analysis of transgenic animals is an integral part of essentially all model organism research. The introduction of fluoresent protein (FP) tags to animal models by Nobel Laureate Martin Chalfie in 1994, using C. elegans, profoundly revolutionized transgenesis by allowing gene expression to be monitored in real time in living organisms. For genes with low level expression, detection of fluorescent protein fusions can be challenging, driving interest in ever more bright and photostable variations of fluorescent proteins. In addition, many applications call for multi-plexing of fluorescently tagged proteins, increasing demand for new proteins with non-overlapping emission spectra. Thus, development of that first green fluorescent protein has inspired discovery and design of numerous variations that can differ in color, brightness, photostability and time to maturation. However, these characteristics can be impacted by environmental conditions such that in vitro properties may not hold in vivo, posing a challenge for researchers looking to expand their FP palette. When a new FP is developed with highly touted characteristics, the in vitro characterized parameters allow selection of good candidates to test in vivo, but adoption of a new protein by the worm community requires significant optimization and in vivo testing. This project will eliminate uncertainties about the in vivo use of new fusion proteins for the C. elegans community by direct quantitative comparison of a set of new FPs relative to each other and to those currently in broad use. Data derived from this project will be used by the C. elegans community broadly to improve experimental designs for diverse research questions, and by researchers using other model organisms to suggest the best candidate FPs to test in vivo in their systems. This project will also result in a set of C. elegans strains expressing FPs for laboratories to test on their own microscopes and/or on demonstration equipment they are considering for purchase. Finally, this project will establish a set of common metrics with which to evaluate future FPs ensuring rigor and reproducibility of testing. All strains and reagents generated in this work will be curated by the CGC and immediately available to the community.