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Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system
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Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system
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Journal Article
Genome editing in the mushroom-forming basidiomycete Coprinopsis cinerea, optimized by a high-throughput transformation system
2017
Overview
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
pro
, 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
pro
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.
Publisher
Nature Publishing Group UK,Nature Portfolio
Subject
