Efficient CRISPR-Cas9 Gene Disruption System in Edible-Medicinal Mushroom .

is a well-known edible medicinal mushroom in East Asia that contains abundant and diverse bioactive compounds. Since traditional genome editing systems in were inefficient and complicated, here, we show that the codon-optimized , which was used with the newly reported promoter Pcmlsm3 and terminator Tcmura3, was expressed. Furthermore, with the help of the negative selection marker , a CRISPR-Cas9 system that included the Cas9 DNA endonuclease, RNA presynthesized and a single-strand DNA template efficiently generated site-specific deletion and insertion. This is the first report of a CRISPR-Cas9 system in , and it could accelerate the genome reconstruction of to meet the need for rapid development in the fungi industry.