A new approach to Cas9-based genome editing in Aspergillus niger that is precise, efficient and selectable.

ABSTRACT: Aspergillus niger and other filamentous fungi are widely used in industry, but efficient genetic engineering of these hosts remains nascent. For example, while molecular genetic tools have been developed, including CRISPR/Cas9, facile genome engineering of A. niger remains challenging. To address these challenges, we have developed a simple Cas9-based gene targeting method that provides selectable, iterative, and ultimately marker-free generation of genomic deletions and insertions. This method leverages locus-specific "pop-out" recombination to suppress off-target integrations. We demonstrated the effectiveness of this method by targeting the phenotypic marker albA and validated it by targeting the glaA and mstC loci. After two selection steps, we observed 100% gene editing efficiency acr

SUBMITTER: Leynaud-Kieffer LMC 

PROVIDER: S-EPMC6336261 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature