Efficient marker free CRISPR/Cas9 genome editing for functional analysis of gene families in filamentous fungi.

ABSTRACT:

Background

CRISPR/Cas9 mediated genome editing has expedited the way of constructing multiple gene alterations in filamentous fungi, whereas traditional methods are time-consuming and can be of mutagenic nature. These developments allow the study of large gene families that contain putatively redundant genes, such as the seven-membered family of crh-genes encoding putative glucan-chitin crosslinking enzymes involved in cell wall biosynthesis.

Results

Here, we present a CRISPR/Cas9 system for Aspergillus niger using a non-integrative plasmid, containing a selection marker, a Cas9 and a sgRNA expression cassette. Combined with selection marker free knockout repair DNA fragments, a set of the seven single knockout strains was obtained through homology directed rep

SUBMITTER: van Leeuwe TM 

PROVIDER: S-EPMC6754632 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature