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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Multiplex genome editing for climate-resilient woody plants

Frontiers in Forests and Global Change, Journal Year: 2025, Volume and Issue: 8

Published: Feb. 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.

Language: Английский

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Unlocking Nature’s Shield: The Promising Potential of CRISPRa in Amplifying Antimicrobial Peptide Expression in Common Bean (Phaseolus vulgaris L.)

ACS Omega, Journal Year: 2025, Volume and Issue: 10(6), P. 5909 - 5918

Published: Feb. 6, 2025

This study proposes using the CRISPR transcriptional activation strategy to modulate expression of genes encoding defense proteins and antimicrobial peptides (AMPs) in Phaseolus vulgaris. Three (PvD1, Pv-thionin, Pv-lectin) were selected targeted by CRISPR-dCas9-TV-mediated complex P. vulgaris L. hairy root. RT-qPCR investigated their efficiency. The eGFP-positive transgenic roots exhibit enhanced compared that control roots. A moderate increase 1.37-fold PvD1 gene was observed roots, while 6.97-fold (Pv-lectin) 5.70-fold (Pv-thionin) increases observed. Importantly, no off-target effects sgRNAs detected, ensuring precision safety CRISPR-dCas9-TV strategy. present article is a proof-of-concept study, it has succeeded demonstrating efficiency modulating target vulgaris, paving way for an alternative approach protecting such essential crop plants.

Language: Английский

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Engineering the auxin-inducible degron system for tunablein vivocontrol of organismal physiology

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 10, 2024

ABSTRACT The physiological mechanisms governing health and disease exhibit complex interactions between multiple genes gene products. To study the dynamics of living systems, researchers need experimental methods capable producing calibrated, quantitative perturbations in vivo — that cannot be obtained using classical genetics, RNAi interference, or small molecule drugs. Recently, an auxin-inducible degron (AID) system has been developed to allow targeted degradation proteins small-molecule activators, providing spatiotemporal control protein abundance. However, a better understanding biochemical activities AID components their context is needed design interventions. Here, we apply engineering approaches characterize understand performance several technologies then improve this multicellular animal Caenorhabditis elegans . We 1) develop new for careful calibration activity specific purposes; 2) TIR1 enzyme constructs with improved over existing constructs; 3) approach simultaneously independently degrade target distinct tissues; finally, 4) pan-organismal by re-engineering enzyme. Taken together, these advances enable cellular systems animals.

Language: Английский

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Towards targeted engineering of promoters via deletion of repressive cis‐regulatory elements

New Phytologist, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 14, 2024

'… this approach is informed by a solid mechanistic understanding of repressive CREs within promoters, presenting more rational and systematic to achieving endogenous gene overexpression.' There are several ways which plant scientists have attempted generate gain-of-function alleles using CRISPR (Fig. 1). The first most widely adopted unbiased, multiplex, CRISPR-mediated, mutagenesis target promoter with multiple guide RNAs (gRNAs) in hope generating via fortuitous upstream regulatory sequences. Since the demonstration Cas9-driven Rodríguez-Leal et al. (2017), groups utilized upregulate their genes interest (Song al., 2022; Zhou 2023; Karavolias 2024; Patel-Tupper 2024). However, method tends result mostly loss-of-function (knock-out knock-down) alleles, some isolated may involve larger-scale rearrangements such as chromosomal inversions (Patel-Tupper Additionally, underlying mechanism driving overexpression frequently not understood approach, limiting its potential for design engineering promoters. A second recently involves insertion enhancers identified genome enrichment or STARR-seq assays followed subsequent introduction enhancer into through CRISPR-mediated knock-in. This one demonstrate overexpression. these elicit also remains elusive. In cases, were either found barely increase expression above levels certain (Claeys 2024) excessive that led stunted growth sterility (Yao third CRISPR-targeted open reading frames (uORFs) present 5′-UTR translation primary ORF (pORF; Zhang 2018). powerful technique increasing protein but cannot be applied lacking uORFs. current study Wang authors introduce fourth overexpress an gene. Unlike previous methods, Recognizing importance improving content major crops, overexpressed NF-YC4, conserved transcription factor known enhance leaf seed (Li 2015), targeted deletion RAV1A WRKY binding motifs from rice soybean NF-YC4 Here, analysis incorporated experimental deserves particular mention: did blindly assume all copies putative repressor-binding site functional. While no has provided satisfying conclusions on how distinguish functional sites (TFBS) nonfunctional ones, many studies highlighted critical role DNA shape flanking sequences conferring affinity specificity plants (Zhang 2019; Sielemann 2021; Li 2023). Notably, (2019) investigating G-box function promoters revealed presence core TFBS motif sufficient function, consequential effects often bordered appropriate sequences, highly motif- context-dependent. Simply deleting possible without obtaining data about elements actually lead desired outcome, resulting lost time effort. used combination orthogonal luciferase reporter vitro electrophoretic mobility shift assay (EMSA) experiments, allowed them focus motifs, thereby facilitating suitable gRNAs CRISPR-based editing efficient manner. It worth noting here successfully transient Nicotiana benthamiana leaves obtain initial readouts driven various variants. Jores (2021) previously limitations studying dicot transcriptional responses monocots, vice versa; however, results indicate well-established model systems N. retain power provide accurate monocot (rice) activity long factors cognate well across monocots dicots. strong correlation between transcript levels, shown expression, strongly supports spare future researchers laborious task isolating protoplasts species validation experiments. Thus, aiming rationally engineer want consider focusing efforts targeting species. By examining carbohydrate different tissues, reveal differences enhancement seeds, raising important questions tissue prompting go tissue-specific future. Another highlight utilization site-directed nuclease (SDN)-1 type edits soybean. three types categorize modification outcomes (Podevin 2013): (1) SDN-1 edits, include double-strand breaks plant's native nonhomologous end joining (NHEJ) repair mechanism; (2) SDN-2 utilize template only few bases differing original sequence; (3) SDN-3 incorporation foreign exogenous piece DNA, typically large considered GM (Ahmad generated deletions simultaneous segments harbouring CREs, presumably repaired NHEJ, so SDN-1. After occurred initially transformed T0 generation, transgene-free recovered generations Mendelian segregation, likely regulated non-GM crops countries according product-based regulations Such simple generalizable accelerate practical impacts research, newly would exempt lengthy approval processes. minimize unintended pleiotropic help preserve regulation. Although tools allow achieve precise interest, need necessarily 100% physiologically relevant desirable phenotype attained. New gene-editing being developed ever-more sequence manipulation, provides roadmap implementing strategy Flora Zhiqi supported part grant number: CZIF2022-007203 Chan Zuckerberg Initiative Foundation. Krishna K. Niyogi investigator at Howard Hughes Medical Institute.

Language: Английский

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Synthetic Genomics in Crop Breeding: Evidence, Opportunities and Challenges

Crop Design, Journal Year: 2024, Volume and Issue: unknown, P. 100090 - 100090

Published: Nov. 1, 2024

Language: Английский

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Recent progress of principal techniques used in the study of Müller glia reprogramming in mice

Cell Regeneration, Journal Year: 2024, Volume and Issue: 13(1)

Published: Dec. 12, 2024

Abstract In zebrafish, Müller glia (MG) cells retain the ability to proliferate and de-differentiate into retinal progenitor-like cells, subsequently differentiating neurons that can replace those damaged or lost due injury. contrast, reprogramming potential of MG in mammals has been lost, with these typically responding damage through gliosis. Considerable efforts have dedicated achieving mammals. Notably, significant advancements achieved mice employing various methodologies. At same time, some inevitable challenges hindered identifying accurate cell rather than illusion, let alone improving efficiency maturity daughter cells. Recently, several strategies, including lineage tracking, multi-omics techniques, functional analysis, developed investigate process mice. This review summarizes both advantages limitations novel strategies for analyzing mice, offering insights enhancing reliability reprogramming. Graphical

Language: Английский

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