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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Protein Engineering

Background:

  • Protein engineering requires screening numerous variants for desired traits.
  • High-throughput creation of protein diversity is achievable, but selection remains difficult.

Purpose of the Study:

  • To develop an integrated screening platform for accelerating protein engineering.
  • To demonstrate the platform's utility by optimizing a far-red fluorescent protein.

Main Methods:

  • A screening platform combining fluorescence-based image analysis and robotic colony picking was constructed.
  • The platform enables tracking of individual colonies and quantitative assessment of library composition.
  • Iterative cycles of mutagenesis and screening were employed.

Main Results:

  • The platform successfully optimized a dim far-red fluorescent protein, increasing its brightness severalfold.
  • A new variant, mCarmine, was developed with enhanced properties for live tissue imaging and molecular tagging.
  • The platform demonstrated flexibility and cost-effectiveness.

Conclusions:

  • The developed screening platform significantly accelerates fluorescence-based protein optimization.
  • mCarmine is a valuable tool for advanced cellular and molecular imaging.
  • The instrumentation offers a powerful, adaptable, and economical solution for protein evolution projects.