CRISPR–Cas applications in agriculture and plant research

Nature Reviews Molecular Cell Biology, Journal Year: 2025, Volume and Issue: unknown

Published: March 7, 2025

Language: Английский

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Intein-mediated split SaCas9 for genome editing in plants

Frontiers in Genome Editing, Journal Year: 2025, Volume and Issue: 6

Published: Jan. 8, 2025

Virus-induced genome editing (VIGE) technologies have been developed to address the limitations plant editing, which heavily relies on genetic transformation and regeneration. However, application of VIGE in plants is hampered by challenge posed size commonly used gene nucleases, Cas9 Cas12a. To overcome this challenge, we employed intein-mediated protein splicing divide SaCas9 transcript into two segments (Split-v1) three (Split-v3). The Split-v1 system demonstrated efficiencies transgenic comparable those achieved with wild-type SaCas9, ranging from 70.2% 96.1%. Additionally, constructed barley stripe mosaic virus (BSMV)-based vectors co-express gRNAs targeting LcHRC, LcGW2, LcTB1 sheepgrass (Leymus chinensis), a Gramineae forage species known for its recalcitrance transformation. Infected leaves exhibited 10.40% 37.03%. These results demonstrate potential split nuclease systems broaden applicability challenging species.

Language: Английский

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Engineering an optimized hypercompact CRISPR/Cas12j‐8 system for efficient genome editing in plants

Plant Biotechnology Journal, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 12, 2025

Summary The Cas12j‐8 nuclease, derived from the type V CRISPR system, is approximately half size of Cas9 and recognizes a 5′‐TTN‐3′ protospacer adjacent motif sequence, thus potentially having broad application in genome editing for crop improvement. However, its efficiency remains low plants. In this study, we rationally engineered both crRNA nuclease. markedly improved When combined, they exhibited robust activity soybean rice, enabling target sites that were previously uneditable. Notably, certain sequences, was comparable to SpCas9 when targeting identical it outperformed Cas12j‐2 variant, nCas12j‐2, across all tested targets. Additionally, developed cytosine base editors based on Cas12j‐8, demonstrating an average increase 5.36‐ 6.85‐fold base‐editing (C T) compared with unengineered system plants, no insertions or deletions (indels) observed. Collectively, these findings indicate hypercompact CRISPR/Cas12j‐8 serves as efficient tool mediated by nuclease cleavage

Language: Английский

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Research Progress and Application of Miniature CRISPR-Cas12 System in Gene Editing

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(23), P. 12686 - 12686

Published: Nov. 26, 2024

CRISPR-Cas system, a natural acquired immune system in prokaryotes that defends against exogenous DNA invasion because of its simple structure and easy operation, has been widely used many research fields such as synthetic biology, crop genetics breeding, precision medicine, so on. The miniature CRISPR-Cas12 an emerging genome editing tool recent years. Compared to the commonly CRISPR-Cas9 CRISPR-Cas12a, unique advantages, rich PAM sites, higher specificity, smaller volume, cytotoxicity. However, application Cas12 proteins methods improve efficiency have not systematically summarized. In this review, we introduce classification summarize structural characteristics type V cleavage mechanism five proteins. gene animals, plants, microorganisms is summarized, strategies are discussed, aiming provide reference for further understanding functional engineering modification system.

Language: Английский

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