Application Note: Automated CRISPR-Based Iterative Genome Engineering for Rapid Strain Improvement

This document describes two applications of the Onyx® Iterative Genome Engineering workflow:

  • In the first, the heterologous green fluorescent protein (GFP) was engineered in the E. coli genome with a library of 723 targeted edits and two rounds of genome engineering, which led to the identification of two novel improved variants in less than 4 months.
  • The second application used a whole-genome approach to improve production of heterologous cellobiohydrolase I (CBH1) enzyme.  Using diverse libraries with >14,000 edits to target both known and unknown genes led to a 4-fold increase in the enzyme production in less than 6 months with 3 rounds of Iterative Genome Engineering.

Download the Application Note

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