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

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...

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Autores principales:	Suyun Moon, Jee Young An, Yeon-Jae Choi, Youn-Lee Oh, Hyeon-Su Ro, Hojin Ryu
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2021
Materias:	crispr/cas9 lentinula edodes hd1 gene editing Botany QK1-989
Acceso en línea:	https://doaj.org/article/dd82cb282f1f469d879beef29d9a354d
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spelling	oai:doaj.org-article:dd82cb282f1f469d879beef29d9a354d2021-12-01T14:40:58ZConstruction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes1229-80932092-932310.1080/12298093.2021.2006401https://doaj.org/article/dd82cb282f1f469d879beef29d9a354d2021-11-01T00:00:00Zhttp://dx.doi.org/10.1080/12298093.2021.2006401https://doaj.org/toc/1229-8093https://doaj.org/toc/2092-9323CRISPR/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 CRISPR-Cas9–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.Suyun MoonJee Young AnYeon-Jae ChoiYoun-Lee OhHyeon-Su RoHojin RyuTaylor & Francis Grouparticlecrispr/cas9lentinula edodeshd1gene editingBotanyQK1-989ENMycobiology, Vol 0, Iss 0, Pp 1-5 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	crispr/cas9 lentinula edodes hd1 gene editing Botany QK1-989
spellingShingle	crispr/cas9 lentinula edodes hd1 gene editing Botany QK1-989 Suyun Moon Jee Young An Yeon-Jae Choi Youn-Lee Oh Hyeon-Su Ro Hojin Ryu Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
description	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 CRISPR-Cas9–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.
format	article
author	Suyun Moon Jee Young An Yeon-Jae Choi Youn-Lee Oh Hyeon-Su Ro Hojin Ryu
author_facet	Suyun Moon Jee Young An Yeon-Jae Choi Youn-Lee Oh Hyeon-Su Ro Hojin Ryu
author_sort	Suyun Moon
title	Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
title_short	Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
title_full	Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
title_fullStr	Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
title_full_unstemmed	Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
title_sort	construction of a crispr/cas9-mediated genome editing system in lentinula edodes
publisher	Taylor & Francis Group
publishDate	2021
url	https://doaj.org/article/dd82cb282f1f469d879beef29d9a354d
work_keys_str_mv	AT suyunmoon constructionofacrisprcas9mediatedgenomeeditingsysteminlentinulaedodes AT jeeyoungan constructionofacrisprcas9mediatedgenomeeditingsysteminlentinulaedodes AT yeonjaechoi constructionofacrisprcas9mediatedgenomeeditingsysteminlentinulaedodes AT younleeoh constructionofacrisprcas9mediatedgenomeeditingsysteminlentinulaedodes AT hyeonsuro constructionofacrisprcas9mediatedgenomeeditingsysteminlentinulaedodes AT hojinryu constructionofacrisprcas9mediatedgenomeeditingsysteminlentinulaedodes
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Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes

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