The CRISPR (clustered regularly interspaced short palindromic repeat)–Cas9 (CRISPR-associated nuclease 9) system is transforming biology by providing a simple and efficient method to precisely edit the genome of any organism. As an important bio industrial workhorse, Aspergillus niger has attracted widespread interest for exploring broaden application in the sectors of industrial enzymes and organic acids. The availability of its genome sequence enables the redesign and engineering novel stains of A. niger based on CRISPR/Cas9 tools. In this study, we have extended CRISPR/Cas9 system for gene editing in A. niger. The endogenous tRNA gene as RNA polymerase III promoter was employed to lead the expression of guide-RNA (gRNA) cassette to delete and replace gene by homologous recombination in Aspergillus niger. The combination of CRISPR/Cas9 system and non-homologous end joining deficient strain allows fast and highly efficiency gene replacement in an organic acids producer A. niger strain with 100 % of transformants showing correct gene replacement.
Full conference title:
- Asperfest 14 (2017)