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

By Yi Zhang, Zhen Liang, Y. Zong, Yanpeng Wang, Jinxing Liu, Kunling Chen, J. Qiu, Caixia Gao. Nature Communications, 2016

791 citations recorded by the source corpus.

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. Plant genome editing typically relies upon transgenic intermediates, which is a concern given the current regulatory requirements concerning GMOs. Here, Zhang et al. describe a method to edit wheat genomes by transiently expressing CRISPR/Cas9 DNA or RNA, and are able to generate mutant plants with no detectable transgenes.

Subfields: Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/2a45503bd2b404ec224bc13163377a62079d8874

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Molecular Biology

1. Jian‐Kang Zhu
2. Nicholas J. Provart
3. M. Davey
4. C. Ridout

In Plant Science

1. Jian‐Kang Zhu
2. Ricardo H. Ramirez-Gonzalez
3. M. Davey
4. Cristobal Uauy
5. C. Ridout
6. Rudi Appels
7. P. Borrill
8. M. Ashraf

Citations in the corpus

• The transcriptional landscape of polyploid wheat (2018, background): “Our work provides a framework for the generation of hypotheses about biological function in polyploid wheat, which can now be experimentally tested using recent developments in sequenced mutant populations (46) and genome editing approaches (47).”
• Genome Editing—Principles and Applications for Functional Genomics Research and Crop Improvement (2017, methodology): “off-target effects are limited in CRISPR–Cas9 edited plants (Zhang et al., 2014; Zhang et al., 2016b).”