Multiplexed, trackable CRISPR-based genome engineering for optimization of heterologous protein expression and activity in E.coli

Advances in CRISPR editing tools have made it possible to engineer proteins and pathways directly in the genome. Editing of heterologous genes in microbial hosts using a stable genomic system provides the advantage of controlled expression that facilitates in-depth examination of sequence-function relationships and investigation of gene expression regulation.

To perform heterologous protein engineering efficiently and cost-effectively, libraries of genetic edits must be designed and generated in multiplex, which necessitates trackability of designs to connect phenotype to genotype. An innovative genome engineering technology – the Onyx™ Digital Genome Engineering Platform – performs genome-wide and trackable CRISPR editing at scale in an automated benchtop device. This approach significantly reduces the time and resources spent on constructing variant libraries, as demonstrated in this Application Note.