Loss of MAR1 Function is a Marker for Co-Selection of CRISPR-Induced Mutations in Plants

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Rinne, J.; Witte, C.-P.; Herde, M.: Loss of MAR1 Function is a Marker for Co-Selection of CRISPR-Induced Mutations in Plants. In: Frontiers in genome editing 3 (2021), 723384. DOI: https://doi.org/10.3389/fgeed.2021.723384

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Sum total of downloads: 84




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Abstract: 
In this study, we describe the establishment of the knockout marker gene MAR1 for selection of CRISPR/Cas9-edited Arabidopsis seedlings and tomato explants in tissue culture. MAR1 encodes a transporter that is located in mitochondria and chloroplasts and is involved in iron homeostasis. It also opportunistically transports aminoglycoside antibiotics into these organelles and defects of the gene render plants insensitive to those compounds. Here, we show that mutations of MAR1 induced by the CRISPR system confer kanamycin-resistance to Arabidopsis plants and tomato tissues. MAR1 is single-copy in a variety of plant species and the corresponding proteins form a distinct phylogenetic clade allowing easy identification of MAR1 orthologs in different plants. We demonstrate that in multiplexing approaches, where Arabidopsis seedlings were selected via a CRISPR/Cas9-induced kanamycin resistance mediated by MAR1 mutation, a mutation in a second target gene was observed with higher frequency than in a control population only selected for the presence of the transgene. This so called co-selection has not been shown before to occur in plants. The technique can be employed to select for edited plants, which might be particularly useful if editing events are rare.
License of this version: CC BY 4.0 Unported
Document Type: Article
Publishing status: publishedVersion
Issue Date: 2021
Appears in Collections:Naturwissenschaftliche Fakultät

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pos. country downloads
total perc.
1 image of flag of Germany Germany 36 42.86%
2 image of flag of United States United States 16 19.05%
3 image of flag of Czech Republic Czech Republic 6 7.14%
4 image of flag of Indonesia Indonesia 5 5.95%
5 image of flag of China China 5 5.95%
6 image of flag of Poland Poland 2 2.38%
7 image of flag of Iran, Islamic Republic of Iran, Islamic Republic of 2 2.38%
8 image of flag of Israel Israel 2 2.38%
9 image of flag of Korea, Republic of Korea, Republic of 1 1.19%
10 image of flag of Japan Japan 1 1.19%
    other countries 8 9.52%

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