High-Efficiency Multiplexed Cytosine Base Editors for Natural Product Synthesis in <i>Yarrowia lipolytica</i>.

ABSTRACT: Yarrowia lipolytica is an industrial host with a high fatty acid flux. Even though CRISPR-based tools have accelerated its metabolic engineering, there remains a need to develop tools for rapid multiplexed strain engineering to accelerate the design-build-test-learn cycle. Base editors have the potential to perform high-efficiency multiplexed gene editing because they do not depend upon double-stranded DNA breaks. Here, we identified that base editors are less toxic than CRISPR-Cas9 for multiplexed gene editing. We increased the editing efficiency by removing the extra nucleotides between tRNA and gRNA and increasing the base editor and gRNA copy number in a Ku70 deficient strain. We achieved five multiplexed gene editing in the ΔKu70 strain at 42% efficiency. Initially, we were uns

SUBMITTER: Ganesan V 

PROVIDER: S-EPMC12478548 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature