Geminiviral-induced genome editing using miniature CRISPR/Cas12j (CasΦ) and Cas12f variants in plants

Gong, Zheng; Previtera, Dominic Andrew; Wang, Yijie; Botella, José Ramón

doi:10.1007/s00299-023-03092-9

Cited by 6 publications

(1 citation statement)

References 10 publications

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“…Cas12a editing has been applied to various crops including rice, wheat, tomato, citrus, soybean and the model plant Arabidopsis thaliana [26] with varied editing efficiencies. Recently miniature variants, CRISPR/Cas12j (CasΦ) and Cas12f systems and were discovered with less than half the size of Cas9 and their orthologs were successfully tested in plants [27]. Efforts are underway to generate improved versions of Cas12a with altered PAM specificities [28; 29] and improved temperature sensitivity for its flexible application and efficient delivery into plant systems [30].…”

Section: Cas12 Proteinmentioning

confidence: 99%

CRISPR/Cas Mediated Genome Engineering in Plants: Application and Future Prospective

Mishra,

Nayak,

Tuteja

et al. 2024

Preprint

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Genetic engineering has become an essential element in developing climate-resilient crops and environmental sustainable solutions to respond to the increasing need for global food security. Genome editing using CRISPR/Cas [Clustered regulatory interspaced short palindromic repeat (CRISPR)-associated protein (Cas)] technology is being applied to a variety of organisms, including plants. This technique has become popular because of its high specificity, effectiveness, and low production cost. Therefore, this technology has the potential to revolutionize agriculture and contribute to global food security. Over the past few years, increasing efforts have been seen in its application in developing higher-yielding, nutrition rich, disease resistant, stress-tolerant “crops” fruits and vegetables. Cas proteins such as Cas9, Cas12, Cas13, and Cas14, among others, have distinct architectures and have been used to create new genetic tools that improve features that are important for agriculture. The versatility of Cas has accelerated genomic analysis and facilitated the use of CRISPR/Cas to manipulate and alter nucleic acid sequences in cells of different organisms. This review provides the evolution of CRISPR technology exploring its mechanisms and contrasting it with traditional breeding and transgenic approaches to improve different stress tolerance. We have also discussed the CRISPR/Cas system and explored three Cas proteins that are currently known to exist: Cas12, Cas13, and Cas14 and their potential to generate foreign DNA free or non-transgenic crops that could be easily regulated for commercialization in most countries.

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Section: Cas12 Proteinmentioning

confidence: 99%

CRISPR/Cas Mediated Genome Engineering in Plants: Application and Future Prospective

Mishra,

Nayak,

Tuteja

et al. 2024

Preprint

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Agrobacterium-Mediated Transformation and Targeted Mutagenesis Using SpCas12f in Rice

Sukegawa,

Toki,

Saika

2024

Methods in Molecular Biology

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Efficient genome editing in rice with miniature Cas12f variants

Ye,

Zhang,

et al. 2024

aBIOTECH

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Genome editing, particularly using the CRISPR/Cas system, has revolutionized biological research and crop improvement. Despite the widespread use of CRISPR/Cas9, it faces limitations such as PAM sequence requirements and challenges in delivering its large protein into plant cells. The hypercompact Cas12f, derived from Acidibacillus sulfuroxidans (AsCas12f), stands out due to its small size of only 422 amino acids and its preference for a T-rich motif, presenting advantageous features over SpCas9. However, its editing efficiency is extremely low in plants. Recent studies have generated two AsCas12f variants, AsCas12f-YHAM and AsCas12f-HKRA, demonstrating higher editing efficiencies in mammalian cells, yet their performance in plants remains unexplored. In this study, through a systematic investigation of genome cleavage activity in rice, we unveiled a substantial enhancement in editing efficiency for both AsCas12f variants, particularly for AsCas12f-HKRA, which achieved an editing efficiency of up to 53%. Furthermore, our analysis revealed that AsCas12f predominantly induces deletion in the target DNA, displaying a unique deletion pattern primarily concentrated at positions 12, 13, 23, and 24, resulting in deletion size mainly of 10 and 11 bp, suggesting significant potential for targeted DNA deletion using AsCas12f. These findings expand the toolbox for efficient genome editing in plants, offering promising prospects for precise genetic modifications in agriculture.

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Geminiviral-induced genome editing using miniature CRISPR/Cas12j (CasΦ) and Cas12f variants in plants

Cited by 6 publications

References 10 publications

CRISPR/Cas Mediated Genome Engineering in Plants: Application and Future Prospective

CRISPR/Cas Mediated Genome Engineering in Plants: Application and Future Prospective

Agrobacterium-Mediated Transformation and Targeted Mutagenesis Using SpCas12f in Rice

Efficient genome editing in rice with miniature Cas12f variants

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