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Exploiting spatial dimensions to enable parallelized continuous directed evolution.

Ting Wei1, Wangsheng Lai1, Qian Chen1,2

  • 1CAS Key Laboratory for Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China.

Molecular Systems Biology
|September 21, 2022
PubMed
Summary
This summary is machine-generated.

We developed SPACE, a simple system for rapid biomolecule evolution using spatial dimensions. This massively parallelizable platform enables directed evolution in general labs, simplifying complex processes.

Keywords:
bacteriophagedirected evolutionrange expansionspatial competitionvirus spreading

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Evolutionary Biology

Background:

  • Current directed evolution technologies often require complex equipment, limiting accessibility.
  • Phage-assisted continuous evolution (PACE) is a powerful but complex system.
  • Bacterial range expansion offers insights into spatial dynamics.

Purpose of the Study:

  • To develop a simplified, highly parallelizable platform for continuous directed evolution.
  • To introduce spatial dimensions into phage-assisted continuous evolution.
  • To create a system applicable to general laboratories.

Main Methods:

  • Developed SPACE (Spatial Phage-assisted Continuous Evolution) by inoculating M13 phages and E. coli on semisolid agar.
  • Leveraged bacterial range expansion and phage-bacteria co-migration.
  • Utilized spatial patterning as an indicator of evolutionary progress.

Main Results:

  • SPACE enables rapid and extensively parallelizable biomolecule evolution.
  • Demonstrated SPACE's ability to evolve T7 RNA polymerase promoter recognition for 96 random sequences in parallel.
  • Showcased simplified experimental setup compared to existing methods.

Conclusions:

  • SPACE is a simple, easily implemented, and massively parallelizable platform for continuous directed evolution.
  • This system broadens the accessibility of directed evolution techniques to general laboratories.
  • SPACE facilitates the emergence of improved or novel protein functions through spatial competition.