Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes

By Zhen Liang, Kunling Chen, Tingdong Li, Yi Zhang, Yanpeng Wang, Qian Zhao, Jinxing Liu, Huawei Zhang, Cuimin Liu, Y. Ran and 1 more. Nature Communications, 2017

798 citations recorded by the source corpus.

Substantial efforts are being made to optimize the CRISPR/Cas9 system for precision crop breeding. The avoidance of transgene integration and reduction of off-target mutations are the most important targets for optimization. Here, we describe an efficient genome editing method for bread wheat using CRISPR/Cas9 ribonucleoproteins (RNPs). Starting from RNP preparation, the whole protocol takes only seven to nine weeks, with four to five independent mutants produced from 100 immature wheat embryos. Deep sequencing reveals that the chance of off-target mutations in wheat cells is much lower in RNP mediated genome editing than in editing with CRISPR/Cas9 DNA. Consistent with this finding, no off-target mutations are detected in the mutant plants. Because no foreign DNA is used in CRISPR/Cas9 RNP mediated genome editing, the mutants obtained are completely transgene free. This method may be widely applicable for producing genome edited crop plants and has a good prospect of being commercialized. Protocols for crop genome editing would ideally be quick, efficient and specific while avoiding integration of transgenes into the genome of edited plants. Here, Lianget al. show that CRISPR/Cas9 ribonucleoproteins can be used to generate genome edited wheat plants in as little as nine weeks.

Subfields: Insect Science, Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/5d97e1d79a219e0e2d649664c07ad7e933d90216

How researchers see this paper

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

In Insect Science

1. Caixia Gao
2. Jian‐Kang Zhu

In Molecular Biology

1. Caixia Gao
2. Kunling Chen
3. Yanpeng Wang
4. Jian‐Kang Zhu
5. H. Raman
6. P. Nicholson

In Plant Science

1. Caixia Gao
2. Kunling Chen
3. Cristobal Uauy
4. Yanpeng Wang
5. Jian‐Kang Zhu
6. P. Nicholson
7. A. Riaz
8. M. Ashraf

Citations in the corpus

• Genome engineering for crop improvement and future agriculture. (2021, positive): “While DNA can be delivered into plant cells by both particle bombardment and Agrobacterium-mediated transformation, RNA and RNP can only be delivered into plant cells by particle bombardment (Liang et al., 2017; Svitashev et al., 2016; Zhang et al., 2016; Figures 2B and 2C).”
• Speed breeding: a powerful tool to accelerate crop research and breeding (2017, background): “Recent advances in genomic tools and resources [19-23] and the decreasing cost of”
• Genome Editing—Principles and Applications for Functional Genomics Research and Crop Improvement (2017, background): “…2015; Wang et al., 2015b; Butler et al., 2016; Forsyth et al., 2016; Jung and Altpeter, 2016; Malnoy et al., 2016; Morineau et al., 2016; Nishitani et al., 2016; Zhang et al., 2016b; Chen et al., 2017; Jia et al., 2017; Jiang et al., 2017; Li et al., 2017a; Liang et al., 2017; Peng et al., 2017).”
• CRISPR/Cas Genome Editing and Precision Plant Breeding in Agriculture. (2019, methodology): “CRISPR/Cas9 RNP delivery by particle bombardment has been reported in maize and wheat (89, 90, 149).”