TY - JOUR T1 - Comparative assessment of fluorescent proteins for <em>in vivo</em> imaging in an animal model system JF - bioRxiv DO - 10.1101/040279 SP - 040279 AU - Jennifer K. Heppert AU - Daniel J. Dickinson AU - Ariel M. Pani AU - Christopher D. Higgins AU - Annette Steward AU - Julie Ahringer AU - Jeffrey R. Kuhn AU - Bob Goldstein Y1 - 2016/01/01 UR - http://biorxiv.org/content/early/2016/02/19/040279.abstract N2 - Fluorescent protein tags are fundamental tools used to visualize gene products and analyze their dynamics in vivo. Recent advances in genome editing have enabled precise insertion of fluorescent protein tags into the genomes of diverse organisms. These advances expand the potential of in vivo imaging experiments, and they facilitate experimentation with new, bright, photostable fluorescent proteins. Most quantitative comparisons of the brightness and photostability of different fluorescent proteins have been made in vitro, removed from biological variables that govern their performance in cells or organisms. To address the gap we quantitatively assessed fluorescent protein properties in vivo in an animal model system. We generated transgenic C. elegans strains expressing green, yellow, or red fluorescent proteins in embryos, and we imaged embryos expressing different fluorescent proteins under the same conditions for direct comparison. We found that mNeonGreen was not bright in vivo as predicted based on in vitro data, but that mNeonGreen is a better tag than GFP for specific kinds of experiments, and we report on optimal red fluorescent proteins. These results identify ideal fluorescent proteins for imaging in vivo in C. elegans embryos, and they suggest good candidate fluorescent proteins to test in other animal model systems.AbbreviationsFPfluorescent proteinGFPgreen fluorescent proteinmNGmonomeric neon greenmYPetmonomeric yellow fluorescent protein for energy transferCRISPRclustered, regularly interspersed, short palindromic repeats ER -