Cas9HF1 enhanced specificity in Ustilago maydis.

Basidiomycota / genetics CRISPR-Associated Protein 9 CRISPR-Cas Systems Clustered Regularly Interspaced Short Palindromic Repeats Gene Editing / methods Genome, Fungal Sensitivity and Specificity Whole Genome Sequencing
CRISPR/Cas9 Fungi genome editing Off-site targets Whole genome sequencing

Journal

Fungal biology
ISSN: 1878-6146
Titre abrégé: Fungal Biol
Pays: Netherlands
ID NLM: 101524465

Informations de publication

Date de publication:
Historique:
received: 18 06 2019
revised: 13 01 2020
accepted: 12 02 2020
entrez: 30 3 2020
pubmed: 30 3 2020
medline: 20 4 2021
Statut: ppublish

Résumé

The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system is widely used as a tool to precisely manipulate genomic sequence targeted by sgRNA (single guide RNA) and is adapted in different species for genome editing. One of the major concerns of CRISPR-Cas9 is the possibility of off-target effects, which can be remedied by the deployment of high fidelity Cas9 variants. Ustilago maydis is a maize fungal pathogen, which has served as a model organism for biotrophic pathogens for decades. The successful adaption of CRISPR-Cas9 in U. maydis greatly facilitated effector biology studies. Here, we constructed an U. maydis reporter strain that allows in vivo quantification of efficiency and target specificity of three high fidelity Cas9 variants, Cas9HF1, Cas9esp1.1 and Cas9hypa. This approach identified Cas9HF1 as most specific Cas9 variant in U. maydis. Furthermore, whole genome sequencing showed absence of off-target effects in U. maydis by CRISPR-Cas9 editing.

Identifiants

DOI: 10.1016/j.funbio.2020.02.006 PMID: 32220383
pubmed: 32220383
pii: S1878-6146(20)30026-X
doi: 10.1016/j.funbio.2020.02.006
pii:
doi:

Substances chimiques

CRISPR-Associated Protein 9 EC 3.1.-

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

228-234

Informations de copyright

Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.

Auteurs

Weiliang Zuo (W)

Botanical Institute and Cluster of Excellence on Plant Sciences (CEPLAS), University of Cologne, BioCenter, Zuelpicher Str. 47a, 50674 Cologne, Germany.

Jasper Rl Depotter (JR)

Botanical Institute and Cluster of Excellence on Plant Sciences (CEPLAS), University of Cologne, BioCenter, Zuelpicher Str. 47a, 50674 Cologne, Germany.

Gunther Doehlemann (G)

Botanical Institute and Cluster of Excellence on Plant Sciences (CEPLAS), University of Cologne, BioCenter, Zuelpicher Str. 47a, 50674 Cologne, Germany. Electronic address: g.doehlemann@uni-koeln.de.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Champignons › Basidiomycota › Cas9HF1 enhanced specificity in Ustilago maydis.
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines associées aux CRISPR › Protéine-9 associée à CRISPR › Cas9HF1 enhanced specificity in Ustilago maydis.
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › Extinction de l'expression des gènes › Systèmes CRISPR-Cas › Cas9HF1 enhanced specificity in Ustilago maydis.
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Génome › Composants de génome › Séquences répétées en tandem › Clustered regularly interspaced short palindromic repeats › Cas9HF1 enhanced specificity in Ustilago maydis.
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Génie génétique › Édition de gène › Cas9HF1 enhanced specificity in Ustilago maydis.
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Génome › Génome microbien › Génome fongique › Cas9HF1 enhanced specificity in Ustilago maydis.
questionsmedicales.fr › Environnement et santé publique › Santé publique › Méthodes épidémiologiques › Statistiques comme sujet › Sensibilité et spécificité › Cas9HF1 enhanced specificity in Ustilago maydis.
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Analyse de séquence › Analyse de séquence d'ADN › Séquençage du génome entier › Cas9HF1 enhanced specificity in Ustilago maydis.