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The HaloTag as a general scaffold for far-red tunable chemigenetic indicators.
Deo, Claire; Abdelfattah, Ahmed S; Bhargava, Hersh K; Berro, Adam J; Falco, Natalie; Farrants, Helen; Moeyaert, Benjamien; Chupanova, Mariam; Lavis, Luke D; Schreiter, Eric R.
Afiliação
  • Deo C; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Abdelfattah AS; Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
  • Bhargava HK; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Berro AJ; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Falco N; Biophysics Graduate Program, University of California, San Francisco, San Francisco, CA, USA.
  • Farrants H; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Moeyaert B; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Chupanova M; Pharmacological Sciences Graduate Program, University of California, Irvine, Irvine, CA, USA.
  • Lavis LD; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
  • Schreiter ER; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA.
Nat Chem Biol ; 17(6): 718-723, 2021 06.
Article em En | MEDLINE | ID: mdl-33795886
Functional imaging using fluorescent indicators has revolutionized biology, but additional sensor scaffolds are needed to access properties such as bright, far-red emission. Here, we introduce a new platform for 'chemigenetic' fluorescent indicators, utilizing the self-labeling HaloTag protein conjugated to environmentally sensitive synthetic fluorophores. We solve a crystal structure of HaloTag bound to a rhodamine dye ligand to guide engineering efforts to modulate the dye environment. We show that fusion of HaloTag with protein sensor domains that undergo conformational changes near the bound dye results in large and rapid changes in fluorescence output. This generalizable approach affords bright, far-red calcium and voltage sensors with highly tunable photophysical and chemical properties, which can reliably detect single action potentials in cultured neurons.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Corantes Fluorescentes / Hidrolases Limite: Animals Idioma: En Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Corantes Fluorescentes / Hidrolases Limite: Animals Idioma: En Ano de publicação: 2021 Tipo de documento: Article