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Video-based pooled screening yields improved far-red genetically encoded voltage indicators.
Tian, He; Davis, Hunter C; Wong-Campos, J David; Park, Pojeong; Fan, Linlin Z; Gmeiner, Benjamin; Begum, Shahinoor; Werley, Christopher A; Borja, Gabriel B; Upadhyay, Hansini; Shah, Himali; Jacques, Jane; Qi, Yitong; Parot, Vicente; Deisseroth, Karl; Cohen, Adam E.
Afiliación
  • Tian H; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
  • Davis HC; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
  • Wong-Campos JD; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
  • Park P; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
  • Fan LZ; Department of Bioengineering, Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA, USA.
  • Gmeiner B; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
  • Begum S; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
  • Werley CA; Q-State Biosciences, Cambridge, MA, USA.
  • Borja GB; Q-State Biosciences, Cambridge, MA, USA.
  • Upadhyay H; Q-State Biosciences, Cambridge, MA, USA.
  • Shah H; Q-State Biosciences, Cambridge, MA, USA.
  • Jacques J; Q-State Biosciences, Cambridge, MA, USA.
  • Qi Y; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
  • Parot V; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
  • Deisseroth K; Institute for Biological and Medical Engineering, Pontificia Universidad Católica de Chile, Santiago, Chile.
  • Cohen AE; Department of Bioengineering, Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA, USA.
Nat Methods ; 20(7): 1082-1094, 2023 07.
Article en En | MEDLINE | ID: mdl-36624211
ABSTRACT
Video-based screening of pooled libraries is a powerful approach for directed evolution of biosensors because it enables selection along multiple dimensions simultaneously from large libraries. Here we develop a screening platform, Photopick, which achieves precise phenotype-activated photoselection over a large field of view (2.3 × 2.3 mm, containing >103 cells, per shot). We used the Photopick platform to evolve archaerhodopsin-derived genetically encoded voltage indicators (GEVIs) with improved signal-to-noise ratio (QuasAr6a) and kinetics (QuasAr6b). These GEVIs gave improved signals in cultured neurons and in live mouse brains. By combining targeted in vivo optogenetic stimulation with high-precision voltage imaging, we characterized inhibitory synaptic coupling between individual cortical NDNF (neuron-derived neurotrophic factor) interneurons, and excitatory electrical synapses between individual hippocampal parvalbumin neurons. The QuasAr6 GEVIs are powerful tools for all-optical electrophysiology and the Photopick approach could be adapted to evolve a broad range of biosensors.
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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Fenómenos Electrofisiológicos / Hipocampo Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Animals Idioma: En Revista: Nat Methods Asunto de la revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: Fenómenos Electrofisiológicos / Hipocampo Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Animals Idioma: En Revista: Nat Methods Asunto de la revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos