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Video-based pooled screening yields improved far-red genetically encoded voltage indicators.

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We developed Photopick, a high-throughput screening platform for biosensor evolution. This platform enabled the development of improved genetically encoded voltage indicators (GEVIs) for advanced neuroscience research.

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Area of Science:

  • Neuroscience
  • Biotechnology
  • Molecular Biology

Background:

  • Directed evolution of biosensors is crucial for advancing biological research.
  • Existing screening methods face limitations in throughput and dimensionality.

Purpose of the Study:

  • To develop a novel screening platform, Photopick, for high-throughput biosensor evolution.
  • To evolve genetically encoded voltage indicators (GEVIs) with enhanced performance using Photopick.

Main Methods:

  • Development of the Photopick platform for precise phenotype-activated photoselection.
  • Utilizing Photopick to evolve archaerhodopsin-derived GEVIs.
  • Application of evolved GEVIs in cultured neurons and live mouse brains.

Main Results:

  • Photopick enables screening of large cell libraries (>10^3 cells) over a 2.3x2.3 mm field of view.
  • Evolved GEVIs (QuasAr6a and QuasAr6b) exhibit improved signal-to-noise ratio and kinetics.
  • GEVIs demonstrated enhanced performance in cultured neurons and in vivo imaging in mouse brains.

Conclusions:

  • The Photopick platform is a powerful tool for directed evolution of biosensors.
  • Evolved QuasAr6 GEVIs are valuable for all-optical electrophysiology.
  • The Photopick approach has broad applicability for evolving diverse biosensors.