Enhancing and expanding the CGC Strain Collection

项目概要/摘要-应用研究部分 隐杆线虫遗传学中心（CGC）旨在提高科学家的效率， 生物医学研究的进展，使用一个首要的模式生物，秀丽隐杆线虫。转基因分析 动物是基本上所有模式生物研究的组成部分。荧光蛋白（FP）的引入 1994年，诺贝尔奖获得者马丁·查尔菲（Martin Chalfie）利用C.优雅的，深刻革命性的 通过允许在活的生物体中真实的时间监测基因表达的转基因。对于基因低 水平表达，荧光蛋白融合的检测可能是具有挑战性的， 和荧光蛋白的光稳定变体。此外，许多应用要求多路复用， 荧光标记的蛋白质，增加了对具有非重叠发射光谱的新蛋白质的需求。因此，在本发明中， 第一个绿色荧光蛋白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不同。然而，这些特征可以 受环境条件的影响，使得体外性质可能无法在体内保持，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， 体外表征的参数允许选择良好的候选物进行体内测试，但采用新的 蠕虫群落对蛋白质的需求需要显著的优化和体内测试。该项目将消除 新的融合蛋白在体内用于C.线虫群落直接定量 一组新的FP相对于彼此和目前广泛使用的FP的比较。由此得出的数据 项目将被C. elegans社区广泛改善实验设计的多样性研究 问题，并由研究人员使用其他模式生物，以建议最佳的候选FP在体内测试， 他们的系统。该项目还将产生一套C。用于实验室测试的表达FP的线虫菌株 他们自己的显微镜和/或他们正在考虑购买的演示设备。最后，这个项目 将建立一套通用指标，用于评估未来的FP，确保 试验.在这项工作中产生的所有菌株和试剂将由CGC管理，并立即提供给 社会各界

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.