Crop Biotech Update

CRISPR Method Successfully Developed for Medicinal Fungus

August 24, 2022
Wild Poria cocos

Scientists from the Hunan University of Chinese Medicine reported the first successful genome editing of Poria cocos using CRISPR-Cas9 system integrating genome-wide off-target prediction and detection. Their findings are expected to support future researchers of the genetic breeding and commercial production of edible and medicinal fungi.

Previous researchers using CRISPR-Cas9 have not identified endogenous RNA polymerase III promoters in P. cocos, hence there is little information known about its gene functional analysis. However, a new study identified the endogenous U6 promoters from the available P. cocos genome data. Scientists used the promoter sequence to construct a sgRNA expression vector Pfc332-PcU6 and developed a protoplast isolation protocol afterward. This allowed them to transform the sgRNA-Cas9 vector into the cells of P. cocos using the PEG/CaCl2-mediated transformation approach and identify off-target sites. They also observed that gene editing efficiency was site-dependent. The scientists successfully disrupted the target marker gene ura3 using the CRISPR-Cas9 system. The findings provide opportunities for genome editing research in medicinal fungus.

Learn more in Frontiers in Microbiology.

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