<named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System

ABSTRACT Clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated gene 9 (CRISPR-Cas9) systems are used for a wide array of genome-editing applications in organisms ranging from fungi to plants and animals. Recently, a CRISPR-Cas9 system has been developed for the dipl...

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Autores principales:	Kyunghun Min, Yuichi Ichikawa, Carol A. Woolford, Aaron P. Mitchell
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Candida albicans genetics Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/ad6ef1cac9904be79eab9291e5684364
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spelling	oai:doaj.org-article:ad6ef1cac9904be79eab9291e56843642021-11-15T15:21:18Z<named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System10.1128/mSphere.00130-162379-5042https://doaj.org/article/ad6ef1cac9904be79eab9291e56843642016-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00130-16https://doaj.org/toc/2379-5042ABSTRACT Clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated gene 9 (CRISPR-Cas9) systems are used for a wide array of genome-editing applications in organisms ranging from fungi to plants and animals. Recently, a CRISPR-Cas9 system has been developed for the diploid fungal pathogen Candida albicans; the system accelerates genetic manipulation dramatically [V. K. Vyas, M. I. Barrasa, and G. R. Fink, Sci Adv 1(3):e1500248, 2015, http://dx.doi.org/10.1126/sciadv.1500248 ]. We show here that the CRISPR-Cas9 genetic elements can function transiently, without stable integration into the genome, to enable the introduction of a gene deletion construct. We describe a transient CRISPR-Cas9 system for efficient gene deletion in C. albicans. Our observations suggest that there are two mechanisms that lead to homozygous deletions: (i) independent recombination of transforming DNA into each allele and (ii) recombination of transforming DNA into one allele, followed by gene conversion of the second allele. Our approach will streamline gene function analysis in C. albicans, and our results indicate that DNA can function transiently after transformation of this organism. IMPORTANCE The fungus Candida albicans is a major pathogen. Genetic analysis of this organism has revealed determinants of pathogenicity, drug resistance, and other unique biological features, as well as the identities of prospective drug targets. The creation of targeted mutations has been greatly accelerated recently through the implementation of CRISPR genome-editing technology by Vyas et al. [Sci Adv 1(3):e1500248, 2015, http://dx.doi.org/10.1126/sciadv.1500248 ]. In this study, we find that CRISPR elements can be expressed from genes that are present only transiently, and we develop a transient CRISPR system that further accelerates C. albicans genetic manipulation.Kyunghun MinYuichi IchikawaCarol A. WoolfordAaron P. MitchellAmerican Society for MicrobiologyarticleCandida albicansgeneticsMicrobiologyQR1-502ENmSphere, Vol 1, Iss 3 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Candida albicans genetics Microbiology QR1-502
spellingShingle	Candida albicans genetics Microbiology QR1-502 Kyunghun Min Yuichi Ichikawa Carol A. Woolford Aaron P. Mitchell <named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System
description	ABSTRACT Clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated gene 9 (CRISPR-Cas9) systems are used for a wide array of genome-editing applications in organisms ranging from fungi to plants and animals. Recently, a CRISPR-Cas9 system has been developed for the diploid fungal pathogen Candida albicans; the system accelerates genetic manipulation dramatically [V. K. Vyas, M. I. Barrasa, and G. R. Fink, Sci Adv 1(3):e1500248, 2015, http://dx.doi.org/10.1126/sciadv.1500248 ]. We show here that the CRISPR-Cas9 genetic elements can function transiently, without stable integration into the genome, to enable the introduction of a gene deletion construct. We describe a transient CRISPR-Cas9 system for efficient gene deletion in C. albicans. Our observations suggest that there are two mechanisms that lead to homozygous deletions: (i) independent recombination of transforming DNA into each allele and (ii) recombination of transforming DNA into one allele, followed by gene conversion of the second allele. Our approach will streamline gene function analysis in C. albicans, and our results indicate that DNA can function transiently after transformation of this organism. IMPORTANCE The fungus Candida albicans is a major pathogen. Genetic analysis of this organism has revealed determinants of pathogenicity, drug resistance, and other unique biological features, as well as the identities of prospective drug targets. The creation of targeted mutations has been greatly accelerated recently through the implementation of CRISPR genome-editing technology by Vyas et al. [Sci Adv 1(3):e1500248, 2015, http://dx.doi.org/10.1126/sciadv.1500248 ]. In this study, we find that CRISPR elements can be expressed from genes that are present only transiently, and we develop a transient CRISPR system that further accelerates C. albicans genetic manipulation.
format	article
author	Kyunghun Min Yuichi Ichikawa Carol A. Woolford Aaron P. Mitchell
author_facet	Kyunghun Min Yuichi Ichikawa Carol A. Woolford Aaron P. Mitchell
author_sort	Kyunghun Min
title	<named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System
title_short	<named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System
title_full	<named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System
title_fullStr	<named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System
title_full_unstemmed	<named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System
title_sort	<named-content content-type="genus-species">candida albicans</named-content> gene deletion with a transient crispr-cas9 system
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/ad6ef1cac9904be79eab9291e5684364
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<named-content content-type="genus-species">Candida albicans</named-content> Gene Deletion with a Transient CRISPR-Cas9 System

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