Targeted DNA ADP-ribosylation triggers templated repair in bacteria and base mutagenesis in eukaryotes.

ABSTRACT: Base editors create precise genomic edits by directing nucleobase deamination or removal without inducing double-stranded DNA breaks. However, a vast chemical space of other DNA modifications remains to be explored for genome editing. Here, we harness the bacterial anti-phage toxin DarT2 to append ADP-ribosyl moieties to DNA, unlocking distinct editing outcomes in bacteria versus eukaryotes. Fusing an attenuated DarT2 to a Cas9 nickase, we program site-specific ADP-ribosylation of thymines within a target DNA sequence. In tested bacteria, targeting drives efficient homologous recombination in tested bacteria, offering flexible and scar-free genome editing without base replacement nor counterselection. In tested eukaryotes including yeast, plants and human cells, targeting drives substituti

SUBMITTER: Patinios C 

PROVIDER: S-EPMC11601458 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature