An efficient CRISPR–Cas12a promoter editing system for crop improvement

Nature Plants, Год журнала: 2023, Номер 9(4), С. 588 - 604

Опубликована: Апрель 6, 2023

Язык: Английский

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CRISPR/Cas-mediated plant genome editing: outstanding challenges a decade after implementation

Trends in Plant Science, Год журнала: 2023, Номер 28(10), С. 1144 - 1165

Опубликована: Июнь 16, 2023

The discovery of the CRISPR/Cas genome-editing system has revolutionized our understanding plant genome. been used for over a decade to modify genomes study specific genes and biosynthetic pathways as well speed up breeding in many species, including both model non-model crops. Although is very efficient genome editing, bottlenecks challenges slow down further improvement applications. In this review we discuss that can occur during tissue culture, transformation, regeneration, mutant detection. We also opportunities provided by new CRISPR platforms applications related gene regulation, abiotic biotic stress response improvement, de novo domestication plants.

Язык: Английский

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Beyond knockouts: fine‐tuning regulation of gene expression in plants with CRISPR‐Cas‐based promoter editing

New Phytologist, Год журнала: 2023, Номер 239(3), С. 868 - 874

Опубликована: Июнь 6, 2023

The CRISPR-Cas-based genome editing field in plants is expanding rapidly. Editing plant promoters to obtain cis-regulatory alleles with altered expression levels or patterns of target genes a highly promising topic. However, primarily used CRISPR-Cas9 has significant limitations when noncoding sequences like promoters, which have unique structures and regulatory mechanisms, including A-T richness, repetitive redundancy, difficulty identifying key regions, higher frequency DNA structure, epigenetic modification, protein binding accessibility issues. Researchers urgently require efficient feasible tools strategies address these obstacles, enhance promoter efficiency, increase diversity polymorphism, and, most importantly, enable 'non-silent' events that achieve precise gene regulation. This article provides insights into the challenges references for implementing editing-based research plants.

Язык: Английский

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Progress and prospect: Biosynthesis of plant natural products based on plant chassis

Biotechnology Advances, Год журнала: 2023, Номер 69, С. 108266 - 108266

Опубликована: Сен. 30, 2023

Язык: Английский

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The type V effectors for CRISPR/Cas-mediated genome engineering in plants

Biotechnology Advances, Год журнала: 2024, Номер 74, С. 108382 - 108382

Опубликована: Май 25, 2024

Язык: Английский

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CRISPR technology towards genome editing of the perennial and semi-perennial crops citrus, coffee and sugarcane

Frontiers in Plant Science, Год журнала: 2024, Номер 14

Опубликована: Янв. 8, 2024

Gene editing technologies have opened up the possibility of manipulating genome any organism in a predicted way. CRISPR technology is most used tool and, agriculture, it has allowed expansion possibilities plant biotechnology, such as gene knockout or knock-in, transcriptional regulation, epigenetic modification, base editing, RNA prime and nucleic acid probing detection. This mostly depends on

Язык: Английский

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High performance TadA-8e derived cytosine and dual base editors with undetectable off-target effects in plants

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Июнь 14, 2024

Abstract Cytosine base editors (CBEs) and adenine (ABEs) enable precise C-to-T A-to-G edits. Recently, ABE8e, derived from TadA-8e, enhances edits in mammalian cells plants. Interestingly, TadA-8e can also be evolved to confer editing. This study compares engineered CBEs rice tomato cells, identifying TadCBEa, TadCBEd, TadCBEd_V106W as efficient with high purity a narrow editing window. A dual editor, TadDE, promotes simultaneous Multiplexed TadCBEa TadDE is demonstrated transgenic rice, no off-target effects detected by whole genome transcriptome sequencing, indicating specificity. Finally, two crop engineering applications using are shown: introducing herbicide resistance alleles OsALS creating synonymous mutations OsSPL14 resist OsMIR156 -mediated degradation. Together, this presents editor valuable additions the plant toolbox.

Язык: Английский

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Expanding plant genome editing scope and profiles with CRISPR‐FrCas9 systems targeting palindromic TA sites

Plant Biotechnology Journal, Год журнала: 2024, Номер 22(9), С. 2488 - 2503

Опубликована: Май 7, 2024

Summary CRISPR‐Cas9 is widely used for genome editing, but its PAM sequence requirements limit efficiency. In this study, we explore Faecalibaculum rodentium Cas9 (FrCas9) plant especially in rice. FrCas9 recognizes a concise 5′‐NNTA‐3′ PAM, targeting more abundant palindromic TA sites genomes than the 5′‐NGG‐3′ of most popular SpCas9. shows cleavage activities at all tested with editing outcomes sharing same characteristics typical system. induces high‐efficiency targeted mutagenesis stable rice lines, readily generating biallelic mutants expected phenotypes. We augment FrCas9's ability to generate larger deletions through fusion exonuclease, TREX2. TREX2‐FrCas9 generates much without compromise demonstrate as an efficient tool genetic knockout microRNA gene. Furthermore, FrCas9‐derived cytosine base editors (CBEs) and adenine (ABE) are developed produce C‐to‐T A‐to‐G edits plants. Whole‐genome sequencing‐based off‐target analysis suggests that highly specific nuclease. Expression plants, however, causes detectable guide RNA‐independent mutations, mostly single nucleotide variants (SNVs). Together, have established CRISPR‐FrCas9 system mutagenesis, large deletions, The simple motif makes promising plants expanded scope.

Язык: Английский

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CRISPR–Cas applications in agriculture and plant research

Nature Reviews Molecular Cell Biology, Год журнала: 2025, Номер unknown

Опубликована: Март 7, 2025

Язык: Английский

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Strategies and Methods for Improving the Efficiency of CRISPR/Cas9 Gene Editing in Plant Molecular Breeding

Plants, Год журнала: 2023, Номер 12(7), С. 1478 - 1478

Опубликована: Март 28, 2023

Following recent developments and refinement, CRISPR-Cas9 gene-editing technology has become increasingly mature is being widely used for crop improvement. The application of CRISPR/Cas9 enables the generation transgene-free genome-edited plants in a short period advantages simplicity, high efficiency, specificity, low production costs, which greatly facilitate study gene functions. In plant molecular breeding, efficiency system proven to be key step influencing effectiveness with improvements recently becoming focus reported scientific research. This review details strategies methods improving editing including Cas9 variant enzyme engineering, effect multiple promoter driven Cas9, gRNA efficient optimization expression strategies. It also briefly introduces CRISPR/Cas12a BE PE precision editing. These are beneficial further development systems field breeding.

Язык: Английский

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