0000000000285773

AUTHOR

Gemma Estrada Girona

showing 3 related works from this author

Fluorogenic Tetrazine-Siliconrhodamine Probe for the Labeling of Noncanonical Amino Acid Tagged Proteins

2018

Tetrazine-bearing fluorescent labels enable site-specific tagging of proteins that are genetically manipulated with dienophile modified noncanonical amino acids. The inverse electron demand Diels-Alder reaction between the tetrazine and the dienophile fulfills the criteria of bioorthogonality allowing fluorescent labeling schemes of live cells. Here, we describe the detailed synthetic and labeling protocols of a near infrared emitting siliconrhodamine-tetrazine probe suitable for super-resolution imaging of residue-specifically engineered proteins in mammalian cells.

0301 basic medicinechemistry.chemical_classificationSuper-resolution microscopy010402 general chemistry01 natural sciencesFluorescence0104 chemical sciencesAmino acid03 medical and health sciencesTetrazinechemistry.chemical_compoundFluorescent labelling030104 developmental biologychemistryBiophysics
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Inducible Genetic Code Expansion in Eukaryotes

2020

Abstract Genetic code expansion (GCE) is a versatile tool to site‐specifically incorporate a noncanonical amino acid (ncAA) into a protein, for example, to perform fluorescent labeling inside living cells. To this end, an orthogonal aminoacyl‐tRNA‐synthetase/tRNA (RS/tRNA) pair is used to insert the ncAA in response to an amber stop codon in the protein of interest. One of the drawbacks of this system is that, in order to achieve maximum efficiency, high levels of the orthogonal tRNA are required, and this could interfere with host cell functionality. To minimize the adverse effects on the host, we have developed an inducible GCE system that enables us to switch on tRNA or RS expression whe…

Context (language use)Computational biology010402 general chemistry01 natural sciencesBiochemistryInsert (molecular biology)Amino Acyl-tRNA SynthetasesRNA TransferEscherichia coliHumansunnatural amino acidAmino AcidsMolecular BiologyT-RExchemistry.chemical_classificationTet-On010405 organic chemistryChemistryCommunicationOrganic ChemistryEukaryotaGenetic codeamber suppressionCommunications0104 chemical sciencesAmino acidMaximum efficiencyFluorescent labellingHEK293 CellsGenetic CodePylRSTransfer RNAMolecular MedicineAmber Stop CodonChemBioChem
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Designer membraneless organelles enable codon reassignment of selected mRNAs in eukaryotes.

2019

How to make an organelle in eukaryotes A key step in the evolution of complex organisms like eukaryotes was the organization of specific tasks into organelles. Reinkemeier et al. designed an artificial, membraneless organelle into mammalian cells to perform orthogonal translation. In response to a specific codon in a selected messenger RNA, ribosomes confined to this organelle were able to introduce chemical functionalities site-specifically, expanding the canonical set of amino acids. This approach opens possibilities in synthetic cell engineering and biomedical research. Science , this issue p. eaaw2644

Computer scienceComputational biology010402 general chemistryProtein Engineering01 natural sciencesGenomeArticle03 medical and health sciencesSynthetic biologyRNA TransferOrganelleChlorocebus aethiopsAnimalsHumansRNA MessengerCaenorhabditis elegansCodon030304 developmental biologyOrganelles0303 health sciencesMultidisciplinaryLysineHEK 293 cellsCell MembraneRNAProtein engineeringGenetic code0104 chemical sciencesHEK293 CellsGenetic CodeProtein BiosynthesisCOS CellsMethanosarcinaSynthetic BiologyRibosomesArtificial OrganellesScience (New York, N.Y.)
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