A promoter assay system using gene targeting in agaricomycetes Pleurotus ostreatus and Coprinopsis cinerea

Dong Xuan Nguyen; Takehito Nakazawa; Genki Myo; Chikako Inoue; Moriyuki Kawauchi; Masahiro Sakamoto; Yoichi Honda

doi:10.1016/j.mimet.2020.106053

A promoter assay system using gene targeting in agaricomycetes Pleurotus ostreatus and Coprinopsis cinerea

J Microbiol Methods. 2020 Dec:179:106053. doi: 10.1016/j.mimet.2020.106053. Epub 2020 Sep 9.

Authors

Dong Xuan Nguyen¹, Takehito Nakazawa², Genki Myo³, Chikako Inoue⁴, Moriyuki Kawauchi⁵, Masahiro Sakamoto⁶, Yoichi Honda⁷

Affiliations

¹ Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan; Biotechnology Center of Ho Chi Minh City, Ho Chi Minh City, Viet Nam. Electronic address: dongmicrobio@gmail.com.
² Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: nakazawa.takehito.8u@kyoto-u.ac.jp.
³ Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: myo.genki.74e@st.kyoto-u.ac.jp.
⁴ Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: inoue.chikako.67m@st.kyoto-u.ac.jp.
⁵ Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: kawauchi.moriyuki.8c@kyoto-u.ac.jp.
⁶ Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: sakamoto.masahiro.6r@kyoto-u.ac.jp.
⁷ Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan. Electronic address: honda.yoichi.5n@kyoto-u.ac.jp.

PMID: 32918936
DOI: 10.1016/j.mimet.2020.106053

Abstract

A novel promoter assay was developed for Agaricomycetes, using a gene-targeting approach, with or without the CRISPR/Cas9 technique. It enables precise evaluation of promoter activity at the original site of the chromosome without random and multiple integrations in conventional transformation experiments.

Keywords: CRISPR/Cas9; Gene targeting; Mushroom; Promoter assay; Wood-rot fungi.

MeSH terms

Agaricales / genetics*
CRISPR-Cas Systems / genetics
Cellulase / genetics
Gene Expression Regulation, Fungal / genetics
Gene Targeting*
Peroxidases / genetics
Pleurotus / genetics*
Promoter Regions, Genetic / genetics*

Substances

Peroxidases
manganese peroxidase
Cellulase

Supplementary concepts

Coprinopsis cinerea