Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA

Abstract Editing plant genomes is technically challenging in hard-to-transform plants and usually involves transgenic intermediates, which causes regulatory concerns. Here we report two simple and efficient genome-editing methods in which plants are regenerated from callus cells transiently expressi...

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Main Authors: Yi Zhang, Zhen Liang, Yuan Zong, Yanpeng Wang, Jinxing Liu, Kunling Chen, Jin-Long Qiu, Caixia Gao
Format: Article
Language:English
Published: Nature Portfolio 2016-08-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/ncomms12617
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author Yi Zhang
Zhen Liang
Yuan Zong
Yanpeng Wang
Jinxing Liu
Kunling Chen
Jin-Long Qiu
Caixia Gao
author_facet Yi Zhang
Zhen Liang
Yuan Zong
Yanpeng Wang
Jinxing Liu
Kunling Chen
Jin-Long Qiu
Caixia Gao
author_sort Yi Zhang
collection DOAJ
description Abstract Editing plant genomes is technically challenging in hard-to-transform plants and usually involves transgenic intermediates, which causes regulatory concerns. Here we report two simple and efficient genome-editing methods in which plants are regenerated from callus cells transiently expressing CRISPR/Cas9 introduced as DNA or RNA. This transient expression-based genome-editing system is highly efficient and specific for producing transgene-free and homozygous wheat mutants in the T0 generation. We demonstrate our protocol to edit genes in hexaploid bread wheat and tetraploid durum wheat, and show that we are able to generate mutants with no detectable transgenes. Our methods may be applicable to other plant species, thus offering the potential to accelerate basic and applied plant genome-engineering research.
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institution OA Journals
issn 2041-1723
language English
publishDate 2016-08-01
publisher Nature Portfolio
record_format Article
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spelling doaj-art-5eeff47d0d90427daddfe48074ff74322025-08-20T01:51:39ZengNature PortfolioNature Communications2041-17232016-08-01711810.1038/ncomms12617Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNAYi Zhang0Zhen Liang1Yuan Zong2Yanpeng Wang3Jinxing Liu4Kunling Chen5Jin-Long Qiu6Caixia Gao7State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesState Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesState Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesState Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesState Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesState Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesState Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of SciencesState Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of SciencesAbstract Editing plant genomes is technically challenging in hard-to-transform plants and usually involves transgenic intermediates, which causes regulatory concerns. Here we report two simple and efficient genome-editing methods in which plants are regenerated from callus cells transiently expressing CRISPR/Cas9 introduced as DNA or RNA. This transient expression-based genome-editing system is highly efficient and specific for producing transgene-free and homozygous wheat mutants in the T0 generation. We demonstrate our protocol to edit genes in hexaploid bread wheat and tetraploid durum wheat, and show that we are able to generate mutants with no detectable transgenes. Our methods may be applicable to other plant species, thus offering the potential to accelerate basic and applied plant genome-engineering research.https://doi.org/10.1038/ncomms12617
spellingShingle Yi Zhang
Zhen Liang
Yuan Zong
Yanpeng Wang
Jinxing Liu
Kunling Chen
Jin-Long Qiu
Caixia Gao
Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA
Nature Communications
title Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA
title_full Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA
title_fullStr Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA
title_full_unstemmed Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA
title_short Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA
title_sort efficient and transgene free genome editing in wheat through transient expression of crispr cas9 dna or rna
url https://doi.org/10.1038/ncomms12617
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