Synonymous edits in the Escherichia coli genome have substantial and condition-dependent effects on fitness.

ABSTRACT: CRISPR-Cas-based genome editing is widely used in bacteria at scales ranging from construction of individual mutants to massively parallel libraries. This procedure relies on guide RNA-directed cleavage of the genome followed by repair with a template that introduces a desired mutation along with synonymous "immunizing" mutations to prevent re-cleavage of the genome after editing. Because the immunizing mutations do not change the protein sequence, they are often assumed to be neutral. However, synonymous mutations can change mRNA structures in ways that alter levels of the encoded proteins. We have tested the assumption that immunizing mutations are neutral by constructing a library of over 50,000 edits that consist of only synonymous mutations in Escherichia coli. Thousands of edi

SUBMITTER: Yang DD 

PROVIDER: S-EPMC10835057 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature