CRISPR technology: A decade of genome editing is only the beginning

Science, Journal Year: 2023, Volume and Issue: 379(6629)

Published: Jan. 19, 2023

The advent of clustered regularly interspaced short palindromic repeat (CRISPR) genome editing, coupled with advances in computing and imaging capabilities, has initiated a new era which genetic diseases individual disease susceptibilities are both predictable actionable. Likewise, genes responsible for plant traits can be identified altered quickly, transforming the pace agricultural research breeding. In this Review, we discuss current state CRISPR-mediated manipulation human cells, animals, plants along relevant successes challenges present roadmap future technology.

Language: Английский

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

Trends in Plant Science, Journal Year: 2023, Volume and Issue: 28(10), P. 1144 - 1165

Published: June 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.

Language: Английский

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Application of CRISPR/Cas9-mediated gene editing for abiotic stress management in crop plants

Frontiers in Plant Science, Journal Year: 2023, Volume and Issue: 14

Published: April 18, 2023

Abiotic stresses, including drought, salinity, cold, heat, and heavy metals, extensively reducing global agricultural production. Traditional breeding approaches transgenic technology have been widely used to mitigate the risks of these environmental stresses. The discovery engineered nucleases as genetic scissors carry out precise manipulation in crop stress-responsive genes associated molecular network has paved way for sustainable management abiotic stress conditions. In this context, clustered regularly interspaced short palindromic repeat-Cas (CRISPR/Cas)-based gene-editing tool revolutionized due its simplicity, accessibility, adaptability, flexibility, wide applicability. This system great potential build up varieties with enhanced tolerance against review, we summarize latest findings on understanding mechanism response plants application CRISPR/Cas-mediated towards a multitude stresses metals. We provide mechanistic insights CRISPR/Cas9-based genome editing technology. also discuss applications evolving techniques such prime base editing, mutant library production, transgene free multiplexing rapidly deliver modern cultivars adapted

Language: Английский

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Advances in Delivery Mechanisms of CRISPR Gene-Editing Reagents in Plants

Frontiers in Genome Editing, Journal Year: 2022, Volume and Issue: 4

Published: Jan. 24, 2022

Gene-editing by CRISPR/Cas systems has revolutionized plant biology serving as a functional genomics tool. It tremendously advanced breeding and crop improvement accelerating the development of improved cultivars, creating genetic variability, aiding in domestication wild orphan crops. is rapidly evolving field. Several advancements include different Cas effectors with increased target range, efficacy, enhanced capacity for precise DNA modifications base editing prime editing. The existing toolbox various CRISPR reagents facilitate gene knockouts, targeted insertions, substitutions, multiplexing. However, major challenge genome-editing remains efficient delivery these into cells. Plants have larger more complex genome structures compared to other living due common occurrence polyploidy re-arrangements. Further, rigid cell walls surrounding cells deter entry any foreign biomolecules. Unfortunately, transformation deliver gene-editing been established only limited number species. Recently, there significant progress plants. This review focuses on exploring mechanisms categorized Agrobacterium -mediated breakthroughs, particle bombardment-based biomolecules recent improvements, protoplasts, versatile system regeneration ultimate goal establish highly genotype-independent reagent multiple targets simultaneously achieve DNA-free gene-edited plants at scale.

Language: Английский

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Highly Efficient Generation of Canker-Resistant Sweet Orange Enabled by an Improved CRISPR/Cas9 System

Frontiers in Plant Science, Journal Year: 2022, Volume and Issue: 12

Published: Jan. 11, 2022

Sweet orange (

Language: Английский

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Green Revolution to Gene Revolution: Technological Advances in Agriculture to Feed the World

Plants, Journal Year: 2022, Volume and Issue: 11(10), P. 1297 - 1297

Published: May 12, 2022

Technological applications in agriculture have evolved substantially to increase crop yields and quality meet global food demand. Conventional techniques, such as seed saving, selective breeding, mutation breeding (variation breeding), dramatically increased production, especially during the 'Green Revolution' 1990s. However, newer issues, limited arable lands, climate change, ever-increasing demand, pose challenges agricultural production threaten security. In following 'Gene era, rapid innovations biotechnology field provide alternative strategies further improve yield, quality, resilience towards biotic abiotic stresses. These include introduction of DNA recombinant technology genome editing transcription activator-like effector (TALEN), zinc-finger nucleases (ZFN), clustered regularly interspaced short palindromic repeats/CRISPR associated (CRISPR/Cas) systems. acceptance future these modern tools rely on regulatory frameworks governing their development various countries. Herein, we examine evolution technological agriculture, focusing motivations for introduction, technical challenges, possible benefits concerns, genetically engineered product production.

Language: Английский

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Sweet Orange: Evolution, Characterization, Varieties, and Breeding Perspectives

Agriculture, Journal Year: 2023, Volume and Issue: 13(2), P. 264 - 264

Published: Jan. 21, 2023

Among Citrus species, the sweet orange (Citrus sinensis (L.) Osbeck) is most important in terms of production volumes and cultivated areas. Oranges are particularly appreciated for organoleptic characteristics high nutraceutical value fruits (thanks especially to their content antioxidants). Recent advances citrus genetic genomic resources, such as release reference genomes several cultivars, have contributed (i) understanding diversification C. its relation with other (ii) assessing molecular mechanisms underlying traits interest, (iii) identifying characterizing candidate genes responsible phenotypic traits, (iv) developing biotechnological methods incorporate these into different genotypes. It has been clarified that all diversity within species was derived from subsequent mutations starting a single ancestor complex cycles hybridization backcrossing between mandarin reticulata Blanco) pummelo maxima (Burm.) Merr.). This paper provides an overview varietal panorama together description main driving forces present future breeding. In fact, orange, well novel varieties improved being pursued thanks employment conventional and/or innovative (molecular-based) methods. The state art innovations genomics tools leading so-called new plant breeding technologies were also reviewed discussed.

Language: Английский

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

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: June 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.

Language: Английский

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Optimized protoplast isolation and transfection with a breakpoint: accelerating Cas9/sgRNA cleavage efficiency validation in monocot and dicot

aBIOTECH, Journal Year: 2024, Volume and Issue: 5(2), P. 151 - 168

Published: April 15, 2024

The CRISPR-Cas genome editing tools are revolutionizing agriculture and basic biology with their simplicity precision ability to modify target genomic loci. Software-predicted guide RNAs (gRNAs) often fail induce efficient cleavage at Many loci inaccessible due complex chromatin structure. Currently, there is no suitable tool available predict the architecture of sites accessibility. Hence, significant time resources spent on performing experiments inefficient guides. Although in vitro-cleavage assay could provide a rough assessment gRNA efficiency, it largely excludes interference native context. Transient in-vivo testing gives proper reagents Here, we developed modified protocol that offers highly protoplast isolation from rice,

Language: Английский

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CRISPR-Cas9 System Mediated Genome Editing Technology: An Ultimate Tool to Enhance Abiotic Stress in Crop Plants

Journal of soil science and plant nutrition, Journal Year: 2024, Volume and Issue: 24(2), P. 1799 - 1822

Published: April 24, 2024

Abstract The drastic rise in the human population globally might uplift issue of food scarcity coming few decades. This problem could affect agricultural sector entirely, and to set targets for uplift, major issues like climate change environmental stresses should be fixed possible high crop production. To develop highly productive resistant varieties using old traditional methods is now a waste time, fast practices use genome editing tools are required. Among all technological tools, CRISPR-Cas9 most precise, productive, quickest system, with extensive usage resist biotic abiotic stresses. technique has direct or indirect influence over quantitative genes withstand shocks. More than 20 crops have been modified CRISPR-Cas improve yield. Researchers CRISPR/Cas-based staple stress resistance improved nutritional quality.Irrespective rules regarding genetically organisms, CRISPR/Cas9 insert through agroinfiltration, viral infection, preassembled Cas9 protein-sgRNA ribonucleoprotein transformation without transgenic impression. Certain undesirable that result starch degradation maltose amassing were deleted by CRISPR reduce cold sensitivity. Precise noxious ion metal removal from roots their effective counterbalancing protoplast notions distant structures also managed gene tools. Spindly knockout creates stress-tolerant (drought salt) plants. can make cost-effective technology multiple sectors. global needs fed as severely affected security, which overcome future advancements CRIPSR technology.

Language: Английский

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