Efficient engineering of chromosomal ribosome binding site libraries in mismatch repair proficient Escherichia coli.

ABSTRACT: Multiplexed gene expression optimization via modulation of gene translation efficiency through ribosome binding site (RBS) engineering is a valuable approach for optimizing artificial properties in bacteria, ranging from genetic circuits to production pathways. Established algorithms design smart RBS-libraries based on a single partially-degenerate sequence that efficiently samples the entire space of translation initiation rates. However, the sequence space that is accessible when integrating the library by CRISPR/Cas9-based genome editing is severely restricted by DNA mismatch repair (MMR) systems. MMR efficiency depends on the type and length of the mismatch and thus effectively removes potential library members from the pool. Rather than working in MMR-deficient strains, which accumula

SUBMITTER: Oesterle S 

PROVIDER: S-EPMC5615074 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature