Efficient genome editing in filamentous fungi via an improved CRISPR‐Cas9 ribonucleoprotein method facilitated by chemical reagents

• Published in: Microbial biotechnology Vol. 14; no. 6; pp. 2343 - 2355
• Main Authors: Zou, Gen, Xiao, Meili, Chai, Shunxing, Zhu, Zhihua, Wang, Ying, Zhou, Zhihua
• Format: Journal Article
• Language: English
• Published: United States John Wiley & Sons, Inc 01.11.2021; John Wiley and Sons Inc; Wiley
• Subjects: Cell division; Cell membranes; Cordyceps - genetics; CRISPR; CRISPR-Cas Systems; Efficiency; Fungi; Fungi - genetics; Gene Editing; Genes; Genetic transformation; Genomes; gRNA; Hypocreales - genetics; Membrane permeability; Mutagenesis; Mutation; Mycelia; Nuclease; Permeability; Plasmids; Polyethylene glycol; Proteins; Protoplasts; Protoplasts - metabolism; Reagents; Regeneration; Ribonucleoproteins - genetics; Ribonucleoproteins - metabolism; RNA, Guide, CRISPR-Cas Systems; Surfactants

We report an optimized genome editing approach for filamentous fungi based on RNPs facilitated by adding chemical reagents. We increased the gene editing efficiency of RNPs significantly by adding Triton X‐100. Reagents related to mitosis and cell division furtherly increased ratio of edited homokar...