Enhancing and expanding the CGC Strain Collection

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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.