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CRISPR/Cas genome editing in soybean: challenges and new insights to overcome existing bottlenecks.

作者信息

Freitas-Alves Nayara Sabrina, Moreira-Pinto Clidia E, Távora Fabiano T P K, Paes-de-Melo Bruno, Arraes Fabricio B M, Lourenço-Tessutti Isabela T, Moura Stéfanie M, Oliveira Antonio C, Morgante Carolina V, Qi Yiping, Fatima Grossi-de-Sa Maria

机构信息

Embrapa Genetic Resources and Biotechnology, Brasília, DF, Brazil; Bioprocess Engineering and Biotechnology Graduate Program, Federal University of Paraná (UFPR), Curitiba, PR, Brazil.

Embrapa Genetic Resources and Biotechnology, Brasília, DF, Brazil; National Institute of Science and Technology, INCT PlantStress Biotech, EMBRAPA, Brasília, DF, Brazil.

出版信息

J Adv Res. 2024 Aug 18. doi: 10.1016/j.jare.2024.08.024.


DOI:10.1016/j.jare.2024.08.024
PMID:39163906
Abstract

BACKGROUND: Soybean is a worldwide-cultivated crop due to its applications in the food, feed, and biodiesel industries. Genome editing in soybean began with ZFN and TALEN technologies; however, CRISPR/Cas has emerged and shortly became the preferable approach for soybean genome manipulation since it is more precise, easy to handle, and cost-effective. Recent reports have focused on the conventional Cas9 nuclease, Cas9 nickase (nCas9) derived base editors, and Cas12a (formally Cpf1) as the most commonly used genome editors in soybean. Nonetheless, several challenges in the complex plant genetic engineering pipeline need to be overcome to effectively edit the genome of an elite soybean cultivar. These challenges include (1) optimizing CRISPR cassette design (i.e., gRNA and Cas promoters, gRNA design and testing, number of gRNAs, and binary vector), (2) improving transformation frequency, (3) increasing the editing efficiency ratio of targeted plant cells, and (4) improving soybean crop production. AIM OF REVIEW: This review provides an overview of soybean genome editing using CRISPR/Cas technology, discusses current challenges, and highlights theoretical (insights) and practical suggestions to overcome the existing bottlenecks. KEY SCIENTIFIC CONCEPTS OF REVIEW: The CRISPR/Cas system was discovered as part of the bacterial innate immune system. It has been used as a biotechnological tool for genome editing and efficiently applied in soybean to unveil gene function, improve agronomic traits such as yield and nutritional grain quality, and enhance biotic and abiotic stress tolerance. To date, the efficiency of gRNAs has been validated using protoplasts and hairy root assays, while stable plant transformation relies on Agrobacterium-mediated and particle bombardment methods. Nevertheless, most steps of the CRISPR/Cas workflow require optimizations to achieve a more effective genome editing in soybean plants.

摘要

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引用本文的文献

[1]
Establishing a CRISPR/Cas9 genome editing framework in pigeonpea (Cajanus cajan L.) by targeting phytoene desaturase (PDS) gene disruption.

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[2]
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[3]
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本文引用的文献

[1]
An ex vitro hairy root system from petioles of detached soybean leaves for in planta screening of target genes and CRISPR strategies associated with nematode bioassays.

Planta. 2023-12-18

[2]
Genetic modification can improve crop yields - but stop overselling it.

Nature. 2023-9

[3]
Discovery of deaminase functions by structure-based protein clustering.

Cell. 2023-7-20

[4]
Boosting genome editing efficiency in human cells and plants with novel LbCas12a variants.

Genome Biol. 2023-4-30

[5]
CRISPR-Cas12a base editors confer efficient multiplexed genome editing in rice.

Plant Commun. 2023-7-10

[6]
Plant Virus-Derived Vectors for Plant Genome Engineering.

Viruses. 2023-2-14

[7]
Systematic optimization of Cas12a base editors in wheat and maize using the ITER platform.

Genome Biol. 2023-1-13

[8]
CRISPR/Cas9-mediated targeted mutation of the decreases photoperiod sensitivity, alters stem growth habits, and decreases branch number in soybean.

Front Plant Sci. 2022-12-14

[9]
Multiplex CRISPR/Cas9-mediated raffinose synthase gene editing reduces raffinose family oligosaccharides in soybean.

Front Plant Sci. 2022-11-15

[10]
The Role of a Soybean 14-3-3 Gene () on White Mold Resistance and Nodulation Investigations Using CRISPR-Cas9 Editing and RNA Silencing.

Mol Plant Microbe Interact. 2023-3

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