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Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes

Summary

This research introduces the first successful genetic editing system for shiitake mushrooms using CRISPR/Cas9 technology. This breakthrough allows scientists to precisely modify shiitake mushroom genes, which has important implications for improving mushroom cultivation and development. Impacts on everyday life: • Could lead to improved shiitake mushroom varieties with better taste, nutrition, and growth characteristics • May help develop mushrooms that are more resistant to diseases and environmental stress • Could enable more efficient and cost-effective mushroom production for food and medicine • May lead to new applications of mushrooms in biotechnology and pharmaceutical production • Could result in more sustainable and environmentally friendly mushroom farming methods

Background

Lentinula edodes (shiitake mushroom) is one of the most widely cultivated edible mushrooms globally, valued for food, seasoning, and pharmacological uses. It shows promise as a cell factory for producing enzymes and organic substances. Despite its importance in food and industrial markets, proper genetic manipulation tools have not been developed for this mushroom species. The CRISPR/Cas9 system has emerged as a powerful genome editing tool used across various organisms but has seen limited application in mushroom-forming fungi.

Objective

To develop and implement the first CRISPR/Cas9 genome editing system in Lentinula edodes by identifying appropriate U6 and GPD promoters from the L. edodes genome and constructing a generic CRISPR/Cas9 plasmid toolkit.

Results

Among the eight tested guide RNAs, three successfully disrupted the LeA1 locus. While CRISPR-Cas9-induced alleles did not affect mating with compatible monokaryotic strains, disruption of transcription levels was detected in the downstream genes of LeHD1 and LeHD2. Two CR-Lehd1 mutants were identified with 2-bp and 33-bp deletion mutations.

Conclusion

The study successfully established the first CRISPR/Cas9 genome editing system in L. edodes, providing a simple and powerful genetic manipulation tool for this scientifically and industrially important edible mushroom. The system demonstrated successful genome editing capability, opening possibilities for future mushroom breeding technologies.
  • Published in:Mycobiology,
  • Study Type:Laboratory Research,
  • Source: 10.1080/12298093.2021.2006401
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