AUTHOR=Chen Bai-Xiong , Wei Tao , Ye Zhi-Wei , Yun Fan , Kang Lin-Zhi , Tang Hong-Biao , Guo Li-Qiong , Lin Jun-Fang 

TITLE=Efficient CRISPR-Cas9 Gene Disruption System in Edible-Medicinal Mushroom Cordyceps militaris

JOURNAL=Frontiers in Microbiology

VOLUME=9

YEAR=2018

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2018.01157

DOI=10.3389/fmicb.2018.01157

ISSN=1664-302X

ABSTRACT=<p><italic>Cordyceps militaris</italic> is a well-known edible medicinal mushroom in East Asia that contains abundant and diverse bioactive compounds. Since traditional genome editing systems in <italic>C. militaris</italic> were inefficient and complicated, here, we show that the codon-optimized <italic>cas9</italic>, which was used with the newly reported promoter Pcmlsm3 and terminator Tcmura3, was expressed. Furthermore, with the help of the negative selection marker <italic>ura3</italic>, a CRISPR-Cas9 system that included the Cas9 DNA endonuclease, RNA presynthesized <italic>in vitro</italic> and a single-strand DNA template efficiently generated site-specific deletion and insertion. This is the first report of a CRISPR-Cas9 system in <italic>C. militaris</italic>, and it could accelerate the genome reconstruction of <italic>C. militaris</italic> to meet the need for rapid development in the fungi industry.</p>