Genetic encoding of a highly photostable, long lifetime fluorescent amino acid for imaging in mammalian cells
Chloe M. Jones, D. Miklos Robkis, Robert J. Blizzard, Mika Munari, Yarra Venkatesh, Tiberiu S. Mihaila, Alex J. Eddins, Ryan A. Mehl, William N. Zagotta, Sharona E. Gordon, E. James Petersson
Abstract
and mammalian cells. We compare protein yields and amino acid specificity for these Acd RSs to identify an optimal construct. We also demonstrate the use of Acd in FLIM, where its long lifetime provides strong contrast compared to endogenous fluorophores and engineered fluorescent proteins, which have lifetimes less than 5 ns.
Topics & Concepts
FluorescenceAmino acidFluorescent proteinMutantQuantum yieldGenetic codeFluorescence-lifetime imaging microscopyGenetic screenChemistryFluorescence microscopeFörster resonance energy transferGreen fluorescent proteinTransfer RNABiophysicsBiochemistryBiologyGeneRNAQuantum mechanicsPhysicsRNA and protein synthesis mechanismsAdvanced Fluorescence Microscopy TechniquesCRISPR and Genetic Engineering