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Tools for genome-wide strain design and construction.

Nanette R Boyle1, Ryan T Gill

  • 1Department of Chemical and Biological Engineering, University of Colorado, Campus Box 424, Boulder, CO 80309, USA.

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Genome-scale directed evolution is now possible using advanced DNA technologies. This enables rapid mapping of mutations and construction of massive combinatorial libraries for biological engineering.

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Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Biotechnology

Background:

  • Directed evolution algorithms have advanced significantly.
  • DNA sequencing and synthesis technologies have improved.
  • Recombinant DNA approaches are increasingly sophisticated.

Purpose of the Study:

  • To review genome-scale directed evolution technologies.
  • To discuss their application in inverse metabolic engineering.
  • To highlight the creation and evaluation of combinatorial libraries.

Main Methods:

  • Genome-scale mapping of mutations to phenotypes.
  • Utilizing advanced DNA sequencing and synthesis.
  • Employing new recombinant DNA approaches.

Main Results:

  • Simultaneous mapping of genome-wide mutations to phenotypes is achievable in under a week.
  • Billions of rationally designed combinatorial mutations can be constructed.
  • Combinatorial libraries can be created and evaluated rapidly.

Conclusions:

  • Genome-scale directed evolution is a powerful new tool.
  • These technologies facilitate inverse metabolic engineering.
  • Rapid library construction and evaluation accelerate biological design.