Genome editing based on <i>in vitro</i>-assembled ribonucleoproteins in plants DOI Creative Commons

Bingyi Liang,

Hikaru Sato, Sachihiro Matsunaga

и другие.

CYTOLOGIA, Год журнала: 2023, Номер 88(4), С. 283 - 288

Опубликована: Дек. 24, 2023

Gene editing technology based on the clustered regularly interspaced short palindromic repeats (CRISPR) system has garnered widespread use in plant genomes because of its cost-effectiveness, efficiency, and simplicity. To avoid integration foreign genes any DNA fragments into target cell genomes, researchers have developed a that introduces vitro-assembled ribonucleoproteins (RNPs) consisting guide RNA (gRNA) Cas protein cells, enabling direct genome editing. This was designed to deliver RNPs through four distinct methods: polyethylene glycol (PEG)-mediated transfection, particle bombardment, electroporation, lipid transfection. In recent years, CRISPR been extensively applied for genetic modification plants, providing strategic response environmental challenges. Researchers successfully established RNP systems various species. Despite some remaining issues, still shows significant promise future applications production non-genetically modified (non-GM) crops.

Язык: Английский

Recent advances of CRISPR-based genome editing for enhancing staple crops DOI Creative Commons
Feng Chen, Chen D. Lu, Yan Zhao

и другие.

Frontiers in Plant Science, Год журнала: 2024, Номер 15

Опубликована: Сен. 23, 2024

An increasing population, climate change, and diminishing natural resources present severe threats to global food security, with traditional breeding genetic engineering methods often falling short in addressing these rapidly evolving challenges. CRISPR/Cas systems have emerged as revolutionary tools for precise modifications crops, offering significant advancements resilience, yield, nutritional value, particularly staple crops like rice maize. This review highlights the transformative potential of technology, emphasizing recent innovations such prime base editing, development novel CRISPR-associated proteins, which significantly improved specificity, efficiency, scope genome editing agriculture. These enable targeted that enhance tolerance abiotic stresses well biotic stresses. Additionally, plays a crucial role improving crop yield quality by enhancing photosynthetic nutrient uptake, resistance lodging, while also taste, texture, shelf life, content through biofortification. Despite challenges off-target effects, need more efficient delivery methods, ethical regulatory concerns, underscores importance security sustainability It calls continued research integration CRISPR other emerging technologies nanotechnology, synthetic biology, machine learning fully realize its developing resilient, productive, sustainable agricultural systems.

Язык: Английский

Процитировано

19

Strategies for delivery of CRISPR/Cas-mediated genome editing to obtain edited plants directly without transgene integration DOI Creative Commons
Zuzana Kocsisova, Viktoriya Coneva

Frontiers in Genome Editing, Год журнала: 2023, Номер 5

Опубликована: Июль 20, 2023

Increased understanding of plant genetics and the development powerful easier-to-use gene editing tools over past century have revolutionized humankind's ability to deliver precise genotypes in crops. Plant transformation techniques are well developed for making transgenic varieties certain crops model organisms, yet reagent delivery regeneration remain key bottlenecks applying technology most Typical protocols produce transgenic, genetically modified (GM) rely on transgenes, chemical selection, tissue culture. make edited (GE) also use even though these may be undesirable final crop product. In some crops, transgenes routinely segregated away during meiosis by performing crosses, thus only a minor concern. other particularly those propagated vegetatively, complex hybrids, or with long generation times, such crosses impractical impossible. This review highlights diverse strategies CRISPR/Cas reagents regenerable cells recover plants without unwanted integration transgenes. Some examples include delivering DNA-free as ribonucleoproteins mRNA, relying expression from non-integrated DNA, using novel mechanisms viruses nanoparticles, unconventional selection methods avoid and/or avoiding culture altogether. These advancing rapidly already enabling scientists precision CRISPR tools.

Язык: Английский

Процитировано

22

Multiplex genome editing for climate-resilient woody plants DOI Creative Commons
Madhab Kumar Sen,

Sunil Kanti Mondal,

Rohit Bharati

и другие.

Frontiers in Forests and Global Change, Год журнала: 2025, Номер 8

Опубликована: Фев. 5, 2025

Climate change is severely impacting global forest ecosystems, stressing woody plants due to rising temperatures, shifting precipitation patterns, and extreme weather events. These pressures threaten biodiversity disrupt the essential roles forests play in carbon sequestration, timber production, ecosystem stability. Traditional management strategies, such as selective breeding, cannot keep up with rapid pace of climate change, given long juvenile phase trees. Multiplex genome editing, particularly through CRISPR technologies, offers a promising solution accelerate development climate-resilient traits plants. By simultaneously targeting multiple genes, multiplex enables efficient modification polygenic that govern stress tolerance, disease resistance, other crucial resilience factors. This mini-review examines potential technologies management, agroecological practices, showing how they can improve tree support sustainable forestry response growing challenges change.

Язык: Английский

Процитировано

0

CRISPR-mediated genome editing of wheat for enhancing disease resistance DOI Creative Commons
Joshua Waites, V. Mohan Murali Achary, Easter D. Syombua

и другие.

Frontiers in Genome Editing, Год журнала: 2025, Номер 7

Опубликована: Фев. 25, 2025

Wheat is cultivated across diverse global environments, and its productivity significantly impacted by various biotic stresses, most importantly but not limited to rust diseases, Fusarium head blight, wheat blast, powdery mildew. The genetic diversity of modern cultivars has been eroded domestication selection, increasing their vulnerability stress due uniformity. rapid spread new highly virulent aggressive pathogen strains exacerbated this situation. Three strategies can be used for enhancing disease resistance through genome editing: introducing (R) gene-mediated resistance, engineering nucleotide-binding leucine-rich repeat receptors (NLRs), manipulating susceptibility (S) genes stop pathogens from exploiting these factors support infection. Utilizing R the common strategy traditional breeding approaches, continuous evolution effectors eventually overcome resistance. Moreover, modifying S confer pleiotropic effects that hinder use in agriculture. Enhancing paramount sustainable production food security, tools are great importance research community. application CRISPR-based editing provides promise improve allowing access a broader range solutions beyond random mutagenesis or intraspecific variation, unlocking ways crops, speeding up breeding. Here, we first summarize major context important diseases limitations. Next, turn our attention powerful applications technology creating varieties against diseases.

Язык: Английский

Процитировано

0

CRISPR plants for studying functional genomics: Methods and applications DOI
Mireia Uranga, Ana Montserrat Martín‐Hernández, Fabio Pasin

и другие.

Elsevier eBooks, Год журнала: 2025, Номер unknown, С. 27 - 59

Опубликована: Янв. 1, 2025

Язык: Английский

Процитировано

0

Characterization Techniques for Nanomaterials in Plant Sciences DOI

T. K. Neelima,

K. M. Fauziya,

B. T. Umesh

и другие.

Опубликована: Янв. 1, 2025

Язык: Английский

Процитировано

0

Microparticle-mediated CRISPR DNA delivery for genome editing in poplar DOI Creative Commons
Lennart Hoengenaert, Jan Van Doorsselaere, Ruben Vanholme

и другие.

Frontiers in Plant Science, Год журнала: 2023, Номер 14

Опубликована: Ноя. 13, 2023

The use of CRISPR/Cas9 is currently the method choice for precise genome engineering in plants, including biomass crop poplar. most commonly used delivering and its components poplar via Agrobacterium-mediated transformation, that besides desired gene-editing event also results stable T-DNA integration. Here we explore delivery reagents DNA-coated microparticle bombardment into model tree Populus tremula x P. alba to evaluate potential developing transgene-free, gene-edited trees, as well integrating donor DNA at specific target sites. Using an optimized transformation method, which favors regeneration plants transiently express genes on delivered DNA, regenerated are free Cas9 antibiotic resistance-encoding transgenes. In addition, report frequent integration fragments Cas9-induced double-strand break, opening opportunities toward targeted gene insertions.

Язык: Английский

Процитировано

8

A new method for the robust expression and single-step purification of dCas9 for CRISPR interference/activation (CRISPRi/a) applications DOI
Harshita Pandey, Binduma Yadav, Koushik Shah

и другие.

Protein Expression and Purification, Год журнала: 2024, Номер 220, С. 106500 - 106500

Опубликована: Май 7, 2024

Язык: Английский

Процитировано

2

Opportunities for Gene Editing of Sweetpotato DOI Creative Commons

Debao Huang,

Chase Livengood,

G. Craig Yencho

и другие.

Compendium of plant genomes, Год журнала: 2024, Номер unknown, С. 123 - 133

Опубликована: Сен. 21, 2024

Язык: Английский

Процитировано

2

Improvement of Gene Delivery and Mutation Efficiency in the CRISPR-Cas9 Wheat (Triticum aestivum L.) Genomics System via Biolistics DOI Open Access
Jaclyn Tanaka, Bastian Minkenberg,

Snigdha Poddar

и другие.

Genes, Год журнала: 2022, Номер 13(7), С. 1180 - 1180

Опубликована: Июнь 30, 2022

Discovery of the CRISPR-Cas9 gene editing system revolutionized field plant genomics. Despite advantages in ease designing gRNA and low cost system, there are still hurdles to overcome mutation efficiencies, specifically hexaploid wheat. In conjunction with delivery transformation frequency, efficiency bottleneck has potential slow down advancements genomic this study, nine bombardment parameter combinations using three gold particle sizes rupture disk pressures were tested establish optimal stable frequencies Utilizing best protocol a knockout cassette phytoene desaturase gene, we subjected transformed embryos four temperature treatments compared efficiencies. The use 0.6 μm particles for increased across all pressures. A heat treatment 34 °C 24 h resulted highest no or minimal reduction frequency. produced two M0 mutant events albino phenotypes, requiring biallelic mutations genomes parameters greatly increases can help advance research efforts wheat

Язык: Английский

Процитировано

9