Base editing enables duplex point mutagenesis in Clostridium autoethanogenum at the price of numerous off-target mutations.

ABSTRACT: Base editors are recent multiplex gene editing tools derived from the Cas9 nuclease of Streptomyces pyogenes. They can target and modify a single nucleotide in the genome without inducing double-strand breaks (DSB) of the DNA helix. As such, they hold great potential for the engineering of microbes that lack effective DSB repair pathways such as homologous recombination (HR) or non-homologous end-joining (NHEJ). However, few applications of base editors have been reported in prokaryotes to date, and their advantages and drawbacks have not been systematically reported. Here, we used the base editors Target-AID and Target-AID-NG to introduce nonsense mutations into four different coding sequences of the industrially relevant Gram-positive bacterium Clostridium autoethanogenum.

SUBMITTER: Seys FM 

PROVIDER: S-EPMC10366002 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature