RT Journal Article SR Electronic T1 Comparative assessment of fluorescent proteins for in vivo imaging in an animal model system JF bioRxiv FD Cold Spring Harbor Laboratory SP 040279 DO 10.1101/040279 A1 Jennifer K. Heppert A1 Daniel J. Dickinson A1 Ariel M. Pani A1 Christopher D. Higgins A1 Annette Steward A1 Julie Ahringer A1 Jeffrey R. Kuhn A1 Bob Goldstein YR 2016 UL http://biorxiv.org/content/early/2016/02/19/040279.abstract AB 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