Genome Editing in the Mushroom-forming Basidiomycete Coprinopsis Cinerea, Optimized by a High-throughput Transformation System

Overview

Journal Sci Rep

Specialty Science

Date 2017 Apr 30

PMID 28455526

Citations 35

Authors

Shigeo S Sugano

Hiroko Suzuki

Eisuke Shimokita

Hirofumi Chiba

Sumihare Noji

Yuriko Osakabe

Keishi Osakabe

Affiliations

Soon will be listed here.

Abstract

Mushroom-forming basidiomycetes produce a wide range of metabolites and have great value not only as food but also as an important global natural resource. Here, we demonstrate CRISPR/Cas9-based genome editing in the model species Coprinopsis cinerea. Using a high-throughput reporter assay with cryopreserved protoplasts, we identified a novel promoter, CcDED1 , with seven times stronger activity in this assay than the conventional promoter GPD2. To develop highly efficient genome editing using CRISPR/Cas9 in C. cinerea, we used the CcDED1 to express Cas9 and a U6-snRNA promoter from C. cinerea to express gRNA. Finally, CRISPR/Cas9-mediated GFP mutagenesis was performed in a stable GFP expression line. Individual genome-edited lines were isolated, and loss of GFP function was detected in hyphae and fruiting body primordia. This novel method of high-throughput CRISPR/Cas9-based genome editing using cryopreserved protoplasts should be a powerful tool in the study of edible mushrooms.

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