Universität zu Köln

Zuo, Weiliang, Depotter, Jasper R. L. and Doehlemann, Gunther ORCID: 0000-0002-7353-8456 (2020). Cas9HF1 enhanced specificity in Ustilago maydis. Fungal Biol., 124 (3-4). S. 228 - 235. OXFORD: ELSEVIER SCI LTD. ISSN 1878-6162

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Abstract

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. (C) 2020 The Authors. Published by Elsevier Ltd on behalf of British Mycological Society.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Zuo, Weiliang UNSPECIFIED UNSPECIFIED UNSPECIFIED Depotter, Jasper R. L. UNSPECIFIED UNSPECIFIED UNSPECIFIED Doehlemann, Gunther UNSPECIFIED orcid.org/0000-0002-7353-8456 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-342173
DOI:	10.1016/j.funbio.2020.02.006
Journal or Publication Title:	Fungal Biol.
Volume:	124
Number:	3-4
Page Range:	S. 228 - 235
Date:	2020
Publisher:	ELSEVIER SCI LTD
Place of Publication:	OXFORD
ISSN:	1878-6162
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Botanical Institute
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language HUMAN-CELLS; U-MAYDIS; RNA; PROTEIN; GENOME; RESISTANCE; DELIVERY Multiple languages Mycology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/34217