Unlocking CRISPR-Cas9 editing for widely diverse Dictyostelid species

ABSTRACT: Dictyostelids are a species-rich clade of cellular slime molds that are widely found in soils and have been studied for over a century. Due to a lack of genome editing methods, most molecular research in Dictyostelids has focused on only a single species, Dictyostelium discoideum, which has severely limited broad-scale comparative analyses. Here, we introduce the first CRISPR-Cas9 editing approach that is cloning-free, selection-free, highly-efficient, and effective across Dictyostelid species that diverged millions of years ago. Depending on the CRISPR-Cas9 target site, our editing approach generates knock-out efficiencies of up to 90% and knock-in efficiencies of up to 50% without selective marker. We show that mutants can be isolated as soon as one day post-transfection, vastly outpacin

SUBMITTER: Mireia Garriga-Canut 

PROVIDER: S-SCDT-10_1038-S44320-025-00180-8 | biostudies-other |

REPOSITORIES: biostudies-other