Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus

Abstract Sporisorium scitamineum is the causative agent responsible for smut disease of sugarcane worldwide. However, lack of efficient gene manipulation system makes this fungus much behind the type model of the smut fungi in molecular biology. Here, we report the development of a CRISPR/Cas9 and T...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Shan Lu, Xiaorui Shen, Baoshan Chen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/eb3dfb9ea57e4a579b54db4c94905f84
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:eb3dfb9ea57e4a579b54db4c94905f84
record_format	dspace
spelling	oai:doaj.org-article:eb3dfb9ea57e4a579b54db4c94905f842021-12-02T11:50:55ZDevelopment of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus10.1038/s41598-017-03233-72045-2322https://doaj.org/article/eb3dfb9ea57e4a579b54db4c94905f842017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-03233-7https://doaj.org/toc/2045-2322Abstract Sporisorium scitamineum is the causative agent responsible for smut disease of sugarcane worldwide. However, lack of efficient gene manipulation system makes this fungus much behind the type model of the smut fungi in molecular biology. Here, we report the development of a CRISPR/Cas9 and T-DNA based dual vector system that allowed efficient knock-out or knock-in of a gene of interest in the S. scitamineum in a site-specific manner. By using Mfa2, a key player in the mating event in S. scitamineum as a tester gene, site-specific insertions of the introduced fragments were achieved both for Mfa2 knockout and complementation. Of particular advantage of this system is the simplicity of selection and identification for the desired transformants by using drug resistance coupled with PCR. This system greatly facilitates the gene function study in S. scitamineum, and could potentially be used for other basidiomycete fungi.Shan LuXiaorui ShenBaoshan ChenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-8 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Shan Lu Xiaorui Shen Baoshan Chen Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus
description	Abstract Sporisorium scitamineum is the causative agent responsible for smut disease of sugarcane worldwide. However, lack of efficient gene manipulation system makes this fungus much behind the type model of the smut fungi in molecular biology. Here, we report the development of a CRISPR/Cas9 and T-DNA based dual vector system that allowed efficient knock-out or knock-in of a gene of interest in the S. scitamineum in a site-specific manner. By using Mfa2, a key player in the mating event in S. scitamineum as a tester gene, site-specific insertions of the introduced fragments were achieved both for Mfa2 knockout and complementation. Of particular advantage of this system is the simplicity of selection and identification for the desired transformants by using drug resistance coupled with PCR. This system greatly facilitates the gene function study in S. scitamineum, and could potentially be used for other basidiomycete fungi.
format	article
author	Shan Lu Xiaorui Shen Baoshan Chen
author_facet	Shan Lu Xiaorui Shen Baoshan Chen
author_sort	Shan Lu
title	Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus
title_short	Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus
title_full	Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus
title_fullStr	Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus
title_full_unstemmed	Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus
title_sort	development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/eb3dfb9ea57e4a579b54db4c94905f84
work_keys_str_mv	AT shanlu developmentofanefficientvectorsystemforgeneknockoutandnearincisgenecomplementationinthesugarcanesmutfungus AT xiaoruishen developmentofanefficientvectorsystemforgeneknockoutandnearincisgenecomplementationinthesugarcanesmutfungus AT baoshanchen developmentofanefficientvectorsystemforgeneknockoutandnearincisgenecomplementationinthesugarcanesmutfungus
_version_	1718395199003033600

Development of an efficient vector system for gene knock-out and near in-cis gene complementation in the sugarcane smut fungus

Ejemplares similares