Author:
Werner Janina,Zuo Weiliang,Doehlemann Gunther
Abstract
AbstractClustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (CRISPR/Cas9) has become the state of art for mutagenesis in filamentous fungi. Here, we describe a RNP-mediated CRISPR/Cas9 for mutagenesis inSporisorium reilianum. The efficiency of the method was testedin vitrowith a cleavage assay as well asin vivowith a GFP-expressingS. reilianumstrain. We applied this method to generate frameshift-, knock-in- and knock-out mutants inS. reilianumwithout a resistance marker by using an auto-replicating plasmid for selection. The RNP-mediated CRISPR/Cas9 increased the mutagenesis efficiency and firstly enables a marker-free genome editing inS. reilianum.
Publisher
Cold Spring Harbor Laboratory
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