A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy.

ABSTRACT: The production of optimized strains of a specific phenotype requires the construction and testing of a large number of genome modifications and combinations thereof. Most bacterial iterative genome-editing methods include essential steps to eliminate selection markers, or to cure plasmids. Additionally, the presence of escapers leads to time-consuming separate single clone picking and subsequent cultivation steps. Herein, we report a genome-editing method based on a Rock-Paper-Scissors (RPS) strategy. Each of three constructed sgRNA plasmids can cure, or be cured by, the other two plasmids in the system; plasmids from a previous round of editing can be cured while the current round of editing takes place. Due to the enhanced curing efficiency and embedded double check mechanism, separate s

SUBMITTER: Wang J 

PROVIDER: S-EPMC7826264 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature