{"doi":"10.1021/jacs.5b06847","title":"Systematic Evaluation of Bioorthogonal Reactions in Live Cells with Clickable HaloTag Ligands: Implications for Intracellular Imaging","abstract":null,"journal":"Journal of the American Chemical Society","year":2015,"id":607538,"datarank":0.7886243058041673,"base_score":5.2574953720277815,"endowment":5.2574953720277815,"self_citation_contribution":0.7886243058041673,"citation_network_contribution":0.0,"self_endowment_contribution":0.7886243058041673,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":191,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1560052,"name":"Joshua C. Judkins","orcid":null,"position":1,"is_corresponding":false},{"id":260571,"name":"Christopher W. am Ende","orcid":"0000-0001-8832-9641","position":2,"is_corresponding":false},{"id":608005,"name":"T. Eric Ballard","orcid":"0000-0003-2511-2810","position":3,"is_corresponding":false},{"id":324005,"name":"Yinzhi Fang","orcid":"0000-0003-1980-0000","position":4,"is_corresponding":false},{"id":1560056,"name":"Keith Riccardi","orcid":null,"position":5,"is_corresponding":false},{"id":451056,"name":"Li Di","orcid":"0000-0001-6117-9022","position":6,"is_corresponding":false},{"id":1560057,"name":"Edward R. Guilmette","orcid":null,"position":7,"is_corresponding":false},{"id":1560058,"name":"Joel W. Schwartz","orcid":null,"position":8,"is_corresponding":false},{"id":324007,"name":"Joseph M. Fox","orcid":"0000-0002-8258-1640","position":9,"is_corresponding":false},{"id":230264,"name":"Douglas S. Johnson","orcid":"0000-0003-2703-6495","position":10,"is_corresponding":false},{"id":1163877,"name":"Heather E. Murrey","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Systematic Evaluation of Bioorthogonal Reactions in Live Cells with Clickable HaloTag Ligands: Implications for Intracellular Imaging","abstract":"Bioorthogonal reactions, including the strain-promoted azide-alkyne cycloaddition (SPAAC) and inverse electron demand Diels-Alder (iEDDA) reactions, have become increasingly popular for live-cell imaging applications. However, the stability and reactivity of reagents has never been systematically explored in the context of a living cell. Here we report a universal, organelle-targetable system based on HaloTag protein technology for directly comparing bioorthogonal reagent reactivity, specificity, and stability using clickable HaloTag ligands in various subcellular compartments. This system enabled a detailed comparison of the bioorthogonal reactions in live cells and informed the selection of optimal reagents and conditions for live-cell imaging studies. We found that the reaction of sTCO with monosubstituted tetrazines is the fastest reaction in cells; however, both reagents have stability issues. To address this, we introduced a new variant of sTCO, Ag-sTCO, which has much improved stability and can be used directly in cells for rapid bioorthogonal reactions with tetrazines. Utilization of Ag complexes of conformationally strained trans-cyclooctenes should greatly expand their usefulness especially when paired with less reactive, more stable tetrazines.","is_dataset_classified":null,"base_score":5.2574953720277815,"endowment":5.2574953720277815,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"26270632","pmcid":"PMC4572613","openalex_id":"https://openalex.org/W2292719107","authors":[],"funders":[{"funder_name":"NIGMS NIH HHS","grant_id":"P30 GM110758","title":null},{"funder_name":"NIBIB NIH HHS","grant_id":"R01EB014354","title":null},{"funder_name":"NCRR NIH HHS","grant_id":"S10RR026962-01","title":null},{"funder_name":"NIGMS NIH HHS","grant_id":"P20 GM104316","title":null},{"funder_name":"NCRR NIH HHS","grant_id":"S10 RR026962","title":null},{"funder_name":"NIH HHS","grant_id":"S10OD016267-01","title":null},{"funder_name":"NIH HHS","grant_id":"S10 OD016267","title":null},{"funder_name":"National Institutes of Health","grant_id":"3P20GM104316-10S1","title":"Discovery of Molecular Probes and Therapeutic Leads"},{"funder_name":"National Institutes of Health","grant_id":"1S10RR026962-01","title":"High Performance Digital NMR Spectrometer"},{"funder_name":"National Institutes of Health","grant_id":"3P20GM104316-07S2","title":"Discovery of Chemical Probes and Therapeutic Leads, Phase II"},{"funder_name":"National Institutes of Health","grant_id":"5P30GM110758-02","title":"Microcopy and Mechanical Testing Core"},{"funder_name":"National Institutes of Health","grant_id":"1S10OD016267-01","title":"Q-Exactive Orbitrap LC-MC/MS System"},{"funder_name":"National Science Foundation","grant_id":"0840401","title":"CRIF:MU Cryprobe Upgrade of a NMR Spectrometer"},{"funder_name":"National Institutes of Health","grant_id":"5R01EB014354-03","title":"The Tetrazine Ligation for Efficient 18F Labeling and Pretargeted Imaging/Radiotherapy of Cancer"},{"funder_name":"National Institutes of Health","grant_id":"5P30GM110758-04","title":"Molecular Design of Advanced Biomaterials"},{"funder_name":"National Science Foundation","grant_id":"1229234","title":"MRI: Acquisition of a High Resolution Mass Spectrometer for Small Molecule Analysis"}],"total_grants":16,"fwci":8.8645,"citation_percentile":0.98457408,"influential_citations":0,"citation_trend":[{"year":2015,"count":1},{"year":2016,"count":20},{"year":2017,"count":19},{"year":2018,"count":14},{"year":2019,"count":15},{"year":2020,"count":18},{"year":2021,"count":29},{"year":2022,"count":11},{"year":2023,"count":25},{"year":2024,"count":17},{"year":2025,"count":17},{"year":2026,"count":4}],"oa_status":"hybrid","license":"other-oa","oa_locations":[{"url":"https://doi.org/10.1021/jacs.5b06847","host_type":"journal"},{"url":"https://doi.org/10.1021/jacs.5b06847","host_type":"publisher"},{"url":"https://pubmed.ncbi.nlm.nih.gov/26270632","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/4572613","host_type":"repository"},{"url":"http://dx.doi.org/10.1021/jacs.5b06847","host_type":""},{"url":"https://dx.doi.org/10.1021/jacs.5b06847","host_type":""}],"fields_of_study":["Click Chemistry and Applications","Monoclonal and Polyclonal Antibodies Research","Advanced Biosensing Techniques and Applications","01 natural sciences","0104 chemical sciences"],"mesh_terms":["Alkenes","Alkynes","Azides","Boron Compounds","Cell Survival","Fluorescent Dyes","HeLa Cells","Humans","Hydrolases","Ligands","Models, Molecular","Protein Conformation","Protein Engineering","Molecular Probes","Fluorescein","Cyclooctanes","Intracellular Space","Molecular Imaging","Cycloaddition Reaction","Hela Cells"],"keywords":["Bioorthogonal chemistry","Chemistry","Reagent","Tetrazine","Cycloaddition","Context (archaeology)","Reactivity (psychology)","Click chemistry","Combinatorial chemistry","Biochemistry","Organic chemistry","Catalysis","Boron Compounds","Models, Molecular","Azides","Cycloaddition Reaction","Cell Survival","Hydrolases","Protein Conformation","Intracellular Space","Alkenes","Ligands","Protein Engineering","Molecular Imaging","Cyclooctanes","Alkynes","Molecular Probes","Humans","Fluorescein","Fluorescent Dyes","HeLa Cells"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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