Establishment of a PEG-mediated protoplast transformation system based on DNA and CRISPR/Cas9 ribonucleoprotein complexes for banana

Wu, Shenping; Zhu, Haocheng; Liu, Jin-Xing; Yang, Qiaosong; Shao, Xiuhong; Bi, Fangcheng; Hu, Chunhua; Huo, Heqiang; Chen, Kunling; Yi, Ganjun

doi:10.1186/s12870-020-02609-8

Cited by 85 publications

(49 citation statements)

References 27 publications

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“…PEG-mediated protoplast assays have been developed for many plant species, including Arabidopsis thaliana , rice ( Oryza sativa ), lettuce ( Lactuca sativa ) ( Woo et al., 2015 ), tobacco ( Nicotiana tabacum and N. attenuata ) ( Woo et al., 2015 ; Kim et al., 2017 ), petunia ( Petunia × hybrida ) ( Subburaj et al., 2016 ; Yu et al., 2020 ), maize ( Zea mays ) ( Sant’Ana et al., 2020 ), grapevine ( Vitis vinifera ), apple ( Malus × domestica ) ( Malnoy et al., 2016 ), wheat ( Triticum aestivum ) ( Liang et al., 2017 ), soybean ( Glycine max ) ( Kim et al., 2017 ; Kim and Choi, 2020 ), potato ( Solanum tuberosum ) ( Andersson et al., 2018 ; González et al., 2020 ), cabbage ( Brassica oleracea ), Chinese cabbage ( Brassica rapa ) ( Murovec et al., 2018 ), and banana ( Musa spp.) ( Wu et al., 2020 ). Up to 71% editing efficiency was achieved in Arabidopsis ( Woo et al., 2015 ).…”

Section: Rnp Delivery Into Plantsmentioning

confidence: 99%

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CRISPR ribonucleoprotein-mediated genetic engineering in plants

Zhang

Iaffaldano

2021

Plant Communications

100

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CRISPR-derived biotechnologies have revolutionized the genetic engineering field and have been widely applied in basic plant research and crop improvement. Commonly used Agrobacterium - or particle bombardment-mediated transformation approaches for the delivery of plasmid-encoded CRISPR reagents can result in the integration of exogenous recombinant DNA and potential off-target mutagenesis. Editing efficiency is also highly dependent on the design of the expression cassette and its genomic insertion site. Genetic engineering using CRISPR ribonucleoproteins (RNPs) has become an attractive approach with many advantages: DNA/transgene-free editing, minimal off-target effects, and reduced toxicity due to the rapid degradation of RNPs and the ability to titrate their dosage while maintaining high editing efficiency. Although RNP-mediated genetic engineering has been demonstrated in many plant species, its editing efficiency remains modest, and its application in many species is limited by difficulties in plant regeneration and selection. In this review, we summarize current developments and challenges in RNP-mediated genetic engineering of plants and provide future research directions to broaden the use of this technology.

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Section: Rnp Delivery Into Plantsmentioning

confidence: 99%

“…Cavendish; AAA Group cv. Baxi) Phytoene desaturase (PDS) Cas9 protoplast PEG targeted deep sequencing 0.19%–0.92% Wu et al. (2020) Wild tobacco ( Nicotiana attenuata ) Phytochrome B, PHYB Cas9 protoplast PEG targeted deep sequencing 44% Woo et al.…”

Section: Rnp Applications In Plant Genome Editingmentioning

confidence: 99%

CRISPR ribonucleoprotein-mediated genetic engineering in plants

Zhang

Iaffaldano

2021

Plant Communications

100

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“…Through the application of these delivery systems, the DNA-free editing approach has been successfully applied to a number of species. A review by Metje-Sprink et al [ 164 ] reports that researchers have achieved transgene-free editing in N. benthaminiana [ 162 ], Solanum tuberosum [ 165 ], T. aestivum , and Z. mays [ 166 , 167 ], Brassicaceae [ 168 ], O. sativa [ 169 ], Musa acuminata [ 170 ], Lactuca sativa [ 171 ], and Piper nigrum [ 172 ]. However, this method has some drawbacks.…”

Section: Challenges and Future Perspectivesmentioning

confidence: 99%

Evolution and Application of Genome Editing Techniques for Achieving Food and Nutritional Security

Fiaz

Ahmar

Saeed

et al. 2021

IJMS

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A world with zero hunger is possible only through a sustainable increase in food production and distribution and the elimination of poverty. Scientific, logistical, and humanitarian approaches must be employed simultaneously to ensure food security, starting with farmers and breeders and extending to policy makers and governments. The current agricultural production system is facing the challenge of sustainably increasing grain quality and yield and enhancing resistance to biotic and abiotic stress under the intensifying pressure of climate change. Under present circumstances, conventional breeding techniques are not sufficient. Innovation in plant breeding is critical in managing agricultural challenges and achieving sustainable crop production. Novel plant breeding techniques, involving a series of developments from genome editing techniques to speed breeding and the integration of omics technology, offer relevant, versatile, cost-effective, and less time-consuming ways of achieving precision in plant breeding. Opportunities to edit agriculturally significant genes now exist as a result of new genome editing techniques. These range from random (physical and chemical mutagens) to non-random meganucleases (MegaN), zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), clustered regularly interspaced short palindromic repeats (CRISPR)/associated protein system 9 (CRISPR/Cas9), the CRISPR system from Prevotella and Francisella1 (Cpf1), base editing (BE), and prime editing (PE). Genome editing techniques that promote crop improvement through hybrid seed production, induced apomixis, and resistance to biotic and abiotic stress are prioritized when selecting for genetic gain in a restricted timeframe. The novel CRISPR-associated protein system 9 variants, namely BE and PE, can generate transgene-free plants with more frequency and are therefore being used for knocking out of genes of interest. We provide a comprehensive review of the evolution of genome editing technologies, especially the application of the third-generation genome editing technologies to achieve various plant breeding objectives within the regulatory regimes adopted by various countries. Future development and the optimization of forward and reverse genetics to achieve food security are evaluated.

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“…Soon after the first successful reports [9], the DNA-free editing approach was applied on a number of species. Since the review of Metje-Sprink et al [8], researchers have achieved DNA-free editing in Nicotiana benthaminiana [10], potato [11], wheat and maize [12,13], Brassicaceae [14], rice [15], banana [16], lettuce [17], pepper [18]. While two years is not a significantly long period, it is clear that commercially important species have become the main target for DNA-free genomic manipulations (i.e.…”

Section: Targeted Plant Species and Delivery Techniques Usedmentioning

confidence: 99%

“…Ribonucleoprotein complexes and nanoparticles are delivered in plant cells mostly by particle bombardment [23] or protoplast transformation. In the latter case lipofection [14,24] or PEG-Ca 2þ transfection [13,15,16] is used. Electroporation is another way to deliver macromolecules across protoplast membrane and well-designed protocols are available [7], but there have been no reports (to our knowledge) during the last two years.…”

Section: Crispr/cas Delivered As Ribonucleoprotein Complexes/nanoparticlesmentioning

confidence: 99%

DNA-free gene editing in plants: a brief overview

Tsanova

Stefanova

Topalova

et al. 2020

Biotechnology & Biotechnological Equipment

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The conversion of bacterial CRISPR/Cas defense system into a simple and efficient tool for genome manipulations brought experimental biology into new dimensions. Suddenly, genome editing reached many groups most of which were interested in it but not able to employ the available time-and labor-consuming approaches of the pre-CRISPR era. In plant biology and biotechnology, CRISPR/Cas gene editing became the second most important technology after plant transformation. Actually, it relies on the available array of methods of gene delivery. While sufficient for most purposes, the classic gene transfer methods might become a problem for some experimental settings. The main obstacle is that they include DNA delivery and, frequently, its subsequent integration into cellular genome. For this reason novel methods to achieve gene editing without the need of stable transformation and even without DNA delivery were developed. These new approaches include in vitro ribonucleoprotein complexes formulations (delivered by microinjection, particle bombardment, electroporation, liposomes etc.), use of virus-like particles and employment of bacterial secretory systems for Cas/gRNA delivery. The first attempts to achieve DNA-free editing were made less than ten years ago. Later, different types of animal and plant cells were addressed. In this mini review we try to summarize the current developments and emerging trends in the field of DNA-free editing in plants.

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Establishment of a PEG-mediated protoplast transformation system based on DNA and CRISPR/Cas9 ribonucleoprotein complexes for banana

Cited by 85 publications

References 27 publications

CRISPR ribonucleoprotein-mediated genetic engineering in plants

CRISPR ribonucleoprotein-mediated genetic engineering in plants

Evolution and Application of Genome Editing Techniques for Achieving Food and Nutritional Security

DNA-free gene editing in plants: a brief overview

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