A robust and practical CRISPR/crRNA screening system for soybean cultivar editing using LbCpf1 ribonucleoproteins

Kim, Hye‐Ran; Choi, Jisun

doi:10.1007/s00299-020-02597-x

Cited by 20 publications

(11 citation statements)

References 29 publications

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“…LbCas12a produced less than 1% editing efficiency in wild tobacco ( N. attenuata ) protoplasts ( Kim et al., 2017 ). In soybean, up to 17.5% editing efficiency was observed in protoplasts using LbCas12a, and the highest editing efficiency obtained using AsCas12a was only 1.6% ( Kim et al., 2017 ; Kim and Choi, 2020 ). Up to 19.3% editing efficiency was observed in pepper ( Capsicum annuum ) using LbCas12a ( Kim et al., 2020 ).…”

Section: Rnp Applications In Plant Genome Editingmentioning

confidence: 98%

“…Efficiencies of different Cas systems and gRNAs can be quickly evaluated using this method. 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 ).…”

Section: Rnp Delivery Into Plantsmentioning

confidence: 99%

“…Williams 82) Fatty acid desaturase 2-1A (FAD2-1A) LbCas12a protoplast PEG targeted deep sequencing 0%–11.7% Kim et al. (2017) Fatty acid desaturase 2-1B (FAD2-1B) 0%–9.1% Fatty acid desaturase 2-1A (FAD2-1A) AsCas12a 0%–1.6% Fatty acid desaturase 2-1B (FAD2-1B) 0%–0.6% Fatty acid desaturase 2-1A (FAD2-1A) LbCas12a cotyledon protoplast PEG targeted deep sequencing 10.5%–11.7% Kim and Choi (2020) Fatty acid desaturase 2-1B (FAD2-1B) 6.7%–9.1% Fatty acid desaturase 2-1A (FAD2-1A) callus protoplast 7.4% Fatty acid desaturase 2-1B (FAD2-1B) 6.9% Soybean ( Glycine max cv. Kwangan) Fatty acid desaturase 2-1A (FAD2-1A) 2.1%–11.8% Fatty acid desaturase 2-1B (FAD2-1B) 1.9%–9.6% Soybean ( Glycine max cv.…”

Section: Rnp Applications In Plant Genome Editingmentioning

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 Applications In Plant Genome Editingmentioning

confidence: 98%

Section: Rnp Delivery Into Plantsmentioning

confidence: 99%

Section: Rnp Applications In Plant Genome Editingmentioning

confidence: 99%

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CRISPR ribonucleoprotein-mediated genetic engineering in plants

Zhang

Iaffaldano

2021

Plant Communications

100

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“…The results showed that the STU is more efficient (Carrijo et al, 2021). Cpf1 (Cas12a) systems have also been established in soybean for GE (Duan et al, 2021;Kim and Choi, 2021). Meanwhile, different GE systems for soybean have been established using specific editing reagent delivery methods developed for soybean transformation, which produce transgene-free GE events either with the biolistic method (Adachi et al, 2021) and selectable marker-free GT systems by O. haywardense H1-8-mediated delivery (Kumar et al, 2022), or by organ-specific editing using an egg cellspecific promoter (Zheng et al, 2020).…”

Section: Development Of Transformation Methods For New Breeding Techn...mentioning

confidence: 99%

“…Transgene-free or DNA-free edited events in many crops can now be obtained either by delivering the RNA form of sgRNA and Cas9 or Cas9 protein (RNP) using the biolistic method, or by protoplast transfection (Chen et al, 2019a;Xu et al, 2020;Gao, 2021;Kim and Choi, 2021). Transgene-free events can also be recovered with the Agrobacterium-mediated method without selection (Liang et al, 2017).…”

Section: Development Of Transformation Methods For New Breeding Techn...mentioning

confidence: 99%

Progress in Soybean Genetic Transformation Over the Last Decade

Guo

Qiu

et al. 2022

Front. Plant Sci.

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Soybean is one of the important food, feed, and biofuel crops in the world. Soybean genome modification by genetic transformation has been carried out for trait improvement for more than 4 decades. However, compared to other major crops such as rice, soybean is still recalcitrant to genetic transformation, and transgenic soybean production has been hampered by limitations such as low transformation efficiency and genotype specificity, and prolonged and tedious protocols. The primary goal in soybean transformation over the last decade is to achieve high efficiency and genotype flexibility. Soybean transformation has been improved by modifying tissue culture conditions such as selection of explant types, adjustment of culture medium components and choice of selection reagents, as well as better understanding the transformation mechanisms of specific approaches such as Agrobacterium infection. Transgenesis-based breeding of soybean varieties with new traits is now possible by development of improved protocols. In this review, we summarize the developments in soybean genetic transformation to date, especially focusing on the progress made using Agrobacterium-mediated methods and biolistic methods over the past decade. We also discuss current challenges and future directions.

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