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BACKGROUND: Pleurotus ostreatus, oyster mushroom, is a popular edible mushroom cultivated world-wide and it also belongs to white-rot fungi that can degrade plant cell wall lignin effectively. To develop an efficient strategy for molecular breeding in P. ostreatus, molecular genetic tools have been developed through my 30-years investigation with wonderful colleagues from domestic and foreign countries. One of the most fruitful and newest international collaborations is that with Korean researchers. In this talk, I would like to introduce some recent results of an international collaboration with Mushroom Division, NIHHS, Rural Development Administration, Republic of Korea. RESULTS: We have successfully introduced a genome editing technology into P. ostreatus using CRISPR/Cas92. To establish a non-genetically modified (non-GM) genome editing in mushroom fungi, we set sail for developing a protocol for CRISPR/Cas9 without foreign DNA integration in the edited strain. In addition, to avoid health and environmental problems caused by spores, we selected and knocked-out candidate genes essential for spore formation using CRISPR/Cas9 technique. Three new genes essential for spore formation were found and sporeless cultivars by disrupting one of these genes were isolated. CONCLUSION: The fundamental technology of non-GM molecular breeding can be used for valuable objectives including high-value-added edible mushrooms, efficient lignin-degrading strains with high potential for bioremediation and non-existent strains su