Transient expression of a TaGRF4-TaGIF1 complex stimulates wheat regeneration and improves genome editing

Qiu, Fengti; Xing, Sinian; Xue, Chenxiao; Liu, Jin-Xing; Chen, Kunling; Chai, Tuanyao; Gao, Caixia

doi:10.1007/s11427-021-1949-9

Cited by 38 publications

(23 citation statements)

References 37 publications

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“…This prompted us to explore novel strategies to enrich these cells. Several DRs have been proved to improve the transformation efficiency by promoting the somatic embryogenesis or regeneration rates ( Lowe et al, 2016 ; Kong et al, 2020 ; Qiu et al, 2022 ). They have been widely used in plants recalcitrant to transformation ( Kong et al, 2020 ; Pan et al, 2022 ; Qiu et al, 2022 ).…”

Section: Resultsmentioning

confidence: 99%

“…Several DRs have been proved to improve the transformation efficiency by promoting the somatic embryogenesis or regeneration rates ( Lowe et al, 2016 ; Kong et al, 2020 ; Qiu et al, 2022 ). They have been widely used in plants recalcitrant to transformation ( Kong et al, 2020 ; Pan et al, 2022 ; Qiu et al, 2022 ). However, the effect of these DRs on the editing efficiency of recipient plants during stable transformation had not yet been investigated.…”

Section: Resultsmentioning

confidence: 99%

“…Several DR ( DEVELPMENTAL REGULATOR ) genes, such as the WUS ( WUSCHEL ), BBM ( BABY BOOM ) and GRFs ( GROWTH-REGULATING FACTORs ), have been proved to improve the transformation efficiency ( Lowe et al, 2016 ; Debernardi et al, 2020 ; Kong et al, 2020 ; Qiu et al, 2022 ). Ectopic expression of the BBM gene, which is originally identified in Brassica napus , has diverse functions in plant cell proliferation, growth and development ( Jha and Kumar, 2018 ).…”

Section: Introductionmentioning

confidence: 99%

“…Several plant-specific GRF transcription factors have successfully elevated the regeneration and transformation efficiency of crop plants, such as soybean, canola, and sunflower ( Kong et al, 2020 ; Pan et al, 2022 ). The overexpression of a chimeric protein consisting of the GRF4 and GRF-interacting factor 1 (GIF1) proteins reinforce the regeneration efficiency and regeneration speed in wheat, triticale, rice and watermelon ( Debernardi et al, 2020 ; Feng et al, 2021 ; Qiu et al, 2022 ). However, the effect of these DRs on the genome editing efficiency in the regenerated plants has not been investigated.…”

Section: Introductionmentioning

confidence: 99%

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Establishment of an Efficient Genome Editing System in Lettuce Without Sacrificing Specificity

Pan

Li²,

Li³

et al. 2022

Front. Plant Sci.

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The efficiency of the CRISPR/Cas9 genome editing system remains limited in many crops. Utilizing strong promoters to boost the expression level of Cas9 are commonly used to improve the editing efficiency. However, these strategies also increase the risk of off-target mutation. Here, we developed a new strategy to utilize intron-mediated enhancement (IME)-assisted 35S promoter to drive Cas9 and sgRNA in a single transcript, which escalates the editing efficiency by moderately enhancing the expression of both Cas9 and sgRNA. In addition, we developed another strategy to enrich cells highly expressing Cas9/sgRNA by co-expressing the developmental regulator gene GRF5, which has been proved to ameliorate the transformation efficiency, and the transgenic plants from these cells also exhibited enhanced editing efficiency. This system elevated the genome editing efficiency from 14–28% to 54–81% on three targets tested in lettuce (Lactuca sativa) without increasing the off-target editing efficiency. Thus, we established a new genome editing system with highly improved on-target editing efficiency and without obvious increasement in off-target effects, which can be used to characterize genes of interest in lettuce and other crops.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Establishment of an Efficient Genome Editing System in Lettuce Without Sacrificing Specificity

Pan

Li²,

Li³

et al. 2022

Front. Plant Sci.

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“…However, strong genotype dependence and highly technical requirements of these systems limit their broad application [ 4 , 5 ]. In recent years, the utilization of developmental regulators (DRs), which are also referred to as morphogenic regulators (MRs), such as the WUS (WUSCHEL), BBM (BABY BOOM) and GRFs (GROWTH-REGULATING FACTORs), have been proven to overcome genotype dependence in plant regeneration [ 6 , 7 , 8 , 9 , 10 , 11 ], while in many plant species, the delivery of exogenous DNA or RNA is genotype dependent and is the main bottleneck in genetic transformation. Therefore, there is an urgent need to develop new delivery vectors for genome engineering.…”

Section: Introductionmentioning

confidence: 99%

The Promising Nanovectors for Gene Delivery in Plant Genome Engineering

Zhi

Zhou

Pan

et al. 2022

IJMS

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Highly efficient gene delivery systems are essential for genetic engineering in plants. Traditional delivery methods have been widely used, such as Agrobacterium-mediated transformation, polyethylene glycol (PEG)-mediated delivery, biolistic particle bombardment, and viral transfection. However, genotype dependence and other drawbacks of these techniques limit the application of genetic engineering, particularly genome editing in many crop plants. There is a great need to develop newer gene delivery vectors or methods. Recently, nanomaterials such as mesoporous silica particles (MSNs), AuNPs, carbon nanotubes (CNTs), and layer double hydroxides (LDHs), have emerged as promising vectors for the delivery of genome engineering tools (DNA, RNA, proteins, and RNPs) to plants in a species-independent manner with high efficiency. Some exciting results have been reported, such as the successful delivery of cargo genes into plants and the generation of genome stable transgenic cotton and maize plants, which have provided some new routines for genome engineering in plants. Thus, in this review, we summarized recent progress in the utilization of nanomaterials for plant genetic transformation and discussed the advantages and limitations of different methods. Furthermore, we emphasized the advantages and potential broad applications of nanomaterials in plant genome editing, which provides guidance for future applications of nanomaterials in plant genetic engineering and crop breeding.

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Gene Editing of Wheat to Reduce Coeliac Disease Epitopes in Gluten

Smulders,

Gilissen,

Juranić

et al. 2023

A Roadmap for Plant Genome Editing

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By using gene editing technologies such as CRISPR/Cas, precise modifications can be made in the genome. CRISPR/Cas is especially valuable for targeted mutagenesis in polyploids, as it can induce mutations of multiple alleles simultaneously, to obtain regenerants that are homozygous for the desired mutation. A range of gene-edited traits have been developed in hexaploid bread wheat, including various nutrition and health-related traits, plant architecture, pest and disease resistance, tolerance to abiotic stress, and traits that enable more efficient breeding. Wheat is also known as a cause of some human diseases, particularly coeliac disease (CD), with a prevalence of 1–2% of the population. In the EU alone, at least 4.5 million people suffer from it. CD is a chronic inflammation of the small intestine, induced and maintained in genetically predisposed individuals by the consumption of gluten proteins from wheat, barley and rye. As there is no cure, patients must follow a life-long gluten-free diet. The dominant epitopes in gluten proteins that trigger the disease, have been characterized, but they cannot be removed by classical breeding without affecting baking quality, as it concerns over 100 gluten genes that occur partly as blocks of genes in the genome of wheat. Using gene editing, two studies have shown that it is possible to modify the epitopes in several alpha- and gamma-gliadins simultaneously, while deleting some of the genes completely. In some lines more than 80% of the alpha-gliadin genes were modified. These proof-of-principle studies show that it is feasible to use gene editing, along with other breeding approaches, to completely remove the CD epitopes from bread wheat. Gene-edited coeliac-safe wheat will have economic, social and environmental impact on food security, nutrition and public health, but the realisation will (partially) depend on new European legislation for plants produced by gene editing.

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Transient expression of a TaGRF4-TaGIF1 complex stimulates wheat regeneration and improves genome editing

Cited by 38 publications

References 37 publications

Establishment of an Efficient Genome Editing System in Lettuce Without Sacrificing Specificity

Establishment of an Efficient Genome Editing System in Lettuce Without Sacrificing Specificity

The Promising Nanovectors for Gene Delivery in Plant Genome Engineering

Gene Editing of Wheat to Reduce Coeliac Disease Epitopes in Gluten

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