Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
10.1080/12298093.2021.2006401
- Author:
Suyun MOON
1
;
Jee Young AN
;
Yeon-Jae CHOI
;
Youn-Lee OH
;
Hyeon-Su RO
;
Hojin RYU
Author Information
1. Department of Biological Sciences and Biotechnology, Chungbuk National University, Cheongju, Korea
- Publication Type:RESEARCH NOTE
- From:Mycobiology
2021;49(6):599-603
- CountryRepublic of Korea
- Language:English
-
Abstract:
CRISPR/Cas9 genome editing systems have been established in a broad range of eukaryotic species. Herein, we report the first method for genetic engineering in pyogo (shiitake) mushrooms (Lentinula edodes) using CRISPR/Cas9. For in vivo expression of guide RNAs (gRNAs) targeting the mating-type gene HD1 (LeA1), we identified an endogenous LeU6 promoter in the L. edodes genome. We constructed a plasmid containing the LeU6 and glyceraldehyde-3-phosphate dehydrogenase (LeGPD) promoters to express the Cas9 protein. Among the eight gRNAs we tested, three successfully disrupted the LeA1 locus. Although the CRISPRCas9–induced alleles did not affect mating with compatible monokaryotic strains, disruption of the transcription levels of the downstream genes of LeHD1 and LeHD2 was detected.Based on this result, we present the first report of a simple and powerful genetic manipulation tool using the CRISPR/Cas9 toolbox for the scientifically and industrially important edible mushroom, L. edodes.
