Gene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas9

Abstract Candida parapsilosis is one of the most common causes of candidiasis, particularly in the very young and the very old. Studies of gene function are limited by the lack of a sexual cycle, the diploid genome, and a paucity of molecular tools. We describe here the development of a plasmid-base...

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Autores principales:	Lisa Lombardi, Siobhán A. Turner, Fang Zhao, Geraldine Butler
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/816dc9fe12bf410f853cbf826e6499f0
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spelling	oai:doaj.org-article:816dc9fe12bf410f853cbf826e6499f02021-12-02T16:07:57ZGene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas910.1038/s41598-017-08500-12045-2322https://doaj.org/article/816dc9fe12bf410f853cbf826e6499f02017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08500-1https://doaj.org/toc/2045-2322Abstract Candida parapsilosis is one of the most common causes of candidiasis, particularly in the very young and the very old. Studies of gene function are limited by the lack of a sexual cycle, the diploid genome, and a paucity of molecular tools. We describe here the development of a plasmid-based CRISPR-Cas9 system for gene editing in C. parapsilosis. A major advantage of the system is that it can be used in any genetic background, which we showed by editing genes in 20 different isolates. Gene editing is carried out in a single transformation step. The CAS9 gene is expressed only when the plasmid is present, and it can be removed easily from transformed strains. There is theoretically no limit to the number of genes that can be edited in any strain. Gene editing is increased by homology-directed repair in the presence of a repair template. Editing by non-homologous end joining (NHEJ) also occurs in some genetic backgrounds. Finally, we used the system to introduce unique tags at edited sites.Lisa LombardiSiobhán A. TurnerFang ZhaoGeraldine ButlerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lisa Lombardi Siobhán A. Turner Fang Zhao Geraldine Butler Gene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas9
description	Abstract Candida parapsilosis is one of the most common causes of candidiasis, particularly in the very young and the very old. Studies of gene function are limited by the lack of a sexual cycle, the diploid genome, and a paucity of molecular tools. We describe here the development of a plasmid-based CRISPR-Cas9 system for gene editing in C. parapsilosis. A major advantage of the system is that it can be used in any genetic background, which we showed by editing genes in 20 different isolates. Gene editing is carried out in a single transformation step. The CAS9 gene is expressed only when the plasmid is present, and it can be removed easily from transformed strains. There is theoretically no limit to the number of genes that can be edited in any strain. Gene editing is increased by homology-directed repair in the presence of a repair template. Editing by non-homologous end joining (NHEJ) also occurs in some genetic backgrounds. Finally, we used the system to introduce unique tags at edited sites.
format	article
author	Lisa Lombardi Siobhán A. Turner Fang Zhao Geraldine Butler
author_facet	Lisa Lombardi Siobhán A. Turner Fang Zhao Geraldine Butler
author_sort	Lisa Lombardi
title	Gene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas9
title_short	Gene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas9
title_full	Gene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas9
title_fullStr	Gene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas9
title_full_unstemmed	Gene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas9
title_sort	gene editing in clinical isolates of candida parapsilosis using crispr/cas9
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/816dc9fe12bf410f853cbf826e6499f0
work_keys_str_mv	AT lisalombardi geneeditinginclinicalisolatesofcandidaparapsilosisusingcrisprcas9 AT siobhanaturner geneeditinginclinicalisolatesofcandidaparapsilosisusingcrisprcas9 AT fangzhao geneeditinginclinicalisolatesofcandidaparapsilosisusingcrisprcas9 AT geraldinebutler geneeditinginclinicalisolatesofcandidaparapsilosisusingcrisprcas9
_version_	1718384667439136768

Gene editing in clinical isolates of Candida parapsilosis using CRISPR/Cas9

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