Development of precise genome editing and multi-copy integration tools in <i>Hansenula polymorpha</i> DL-1.

ABSTRACT: Hansenula polymorpha DL-1 is a thermotolerant yeast capable of utilizing multiple renewable carbon sources, making it a promising microbial cell factory for sustainable manufacturing. However, advanced metabolic engineering efforts have been constrained by its strong non-homologous end joining (NHEJ) mechanism and limited choice of suitable genetic tools. This study presents an optimized synthetic biology toolkit to address these limitations. A high-efficiency CRISPR-Cas9-based genome editing system was established, achieving an editing efficiency of 97.2 %. To further enhance homologous recombination (HR), the NHEJ pathway was partially suppressed by knocking out KU80 and overexpressing HR-related genes from Saccharomyces cerevisiae. This increased HR rates to 88.9 %.

SUBMITTER: Zou X 

PROVIDER: S-EPMC12305584 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature