Current Advancement and Future Prospects in Simplified Transformation-Based Plant Genome Editing DOI Creative Commons
Xueying Han, Ziniu Deng, Huiyun Liu

et al.

Plants, Journal Year: 2025, Volume and Issue: 14(6), P. 889 - 889

Published: March 12, 2025

Recent years have witnessed remarkable progress in plant biology, driven largely by the rapid evolution of CRISPR/Cas-based genome editing (GE) technologies. These tools, including versatile CRISPR/Cas systems and their derivatives, such as base editors prime editors, significantly enhanced universality, efficiency, convenience functional genomics, genetics, molecular breeding. However, traditional genetic transformation methods are essential for obtaining GE plants. depend on tissue culture procedures, which time-consuming, labor-intensive, genotype-dependent, challenging to regenerate. Here, we systematically outline current advancements simplifying GE, focusing optimization process through developmental regulators, development planta methods, establishment nanomaterial- viral vector-based delivery platforms. We also discuss critical challenges future directions achieving genotype-independent, culture-free GE.

Language: Английский

Exploiting viral vectors to deliver genome editing reagents in plants DOI Creative Commons

Yilin Shen,

Ye Tao, Zihan Li

et al.

aBIOTECH, Journal Year: 2024, Volume and Issue: 5(2), P. 247 - 261

Published: May 8, 2024

Genome editing holds great promise for the molecular breeding of plants, yet its application is hindered by shortage simple and effective means delivering genome reagents into plants. Conventional plant transformation-based methods delivery plants often involve prolonged tissue culture, a labor-intensive technically challenging process many elite crop cultivars. In this review, we describe various virus-based that have been employed to deliver reagents, including components CRISPR/Cas machinery donor DNA precision in We update progress these with recent successful examples achieved through different species, highlight advantages limitations approaches, discuss remaining challenges.

Language: Английский

Citations

7
CRISPR/Cas-mediated germplasm improvement and new strategies for crop protection DOI Creative Commons
Ganggang Dong, Zaifeng Fan

Crop Health, Journal Year: 2024, Volume and Issue: 2(1)

Published: Feb. 1, 2024

Abstract Global agriculture and food security are encountering unprecedented challenges from both the ever-growing population rapidly changing climate conditions. CRISPR/Cas-mediated genome editing technology has revolutionized plant functional genetic research precision crop breeding with robustness, high target specificity programmability. Furthermore, numerous emerging biotechnologies based on CRISPR/Cas platform provide opportunity to create new germplasms durable resistance against disease or insect pests, herbicide tolerance, other stress-tolerant improvements, reshaping protection increase agricultural resilience sustainability. In this review, we briefly describe toolbox, including base editing, prime compact manipulation, transcriptional regulation epigenetic then overview most important applications of improvement, highlighting protection-based stress engineering. addition, enumerate global regulations genome-edited crops. Finally, discuss some bottlenecks facing cutting-edge infinite possibilities for future.

Language: Английский

Citations

6
Efficient and transformation‐free genome editing in pepper enabled by RNA virus‐mediated delivery of CRISPR/Cas9 DOI

Chenglu Zhao,

Huanhuan Lou,

Qian Liu

et al.

Journal of Integrative Plant Biology, Journal Year: 2024, Volume and Issue: 66(10), P. 2079 - 2082

Published: July 10, 2024

Tomato spotted wilt virus-mediated delivery of CRISPR/Cas9 bypasses the need for stable transformation and permits efficient, DNA-free genome editing in pepper. Remarkably, up to 77.9% regenerated pepper plants contained heritable edits. This method has been validated with two varieties is compatible existing tissue culture protocols.

Language: Английский

Citations

6
Development of virus-induced genome editing methods in Solanaceous crops DOI Creative Commons
Seoyoung Lee,

Bomi Kang,

Jelli Venkatesh

et al.

Horticulture Research, Journal Year: 2023, Volume and Issue: 11(1)

Published: Nov. 17, 2023

ABSTRACT Genome editing (GE) using CRISPR/Cas systems has revolutionized plant mutagenesis. However, conventional transgene-mediated GE methods have limitations due to the time-consuming generation of stable transgenic lines expressing Cas9/single guide RNA (sgRNA) module through tissue cultures. Virus-induced genome (VIGE) been successfully employed in model plants, such as Arabidopsis thaliana and Nicotiana spp. In this study, we developed two VIGE for Solanaceous plants. First, used tobacco rattle virus (TRV) vector deliver sgRNAs into a tomato (Solanum lycopersicum) line cultivar Micro-Tom Cas9. Second, devised transgene-free method based on potato X (PVX) Cas9 sgRNAs. We designed cloned targeting Phytoene desaturase vectors determined optimal conditions VIGE. evaluated efficiency deep sequencing target gene after viral inoculation, detecting 40.3% 36.5% mutation rates TRV- PVX-mediated GE, respectively. To improve efficiency, applied 37°C heat treatment, which increased by 33% 46% 56% 76% VIGE, obtain edited subjected inoculated cotyledons culture, yielding successful events. also demonstrated that can be other crops, tuberosum) eggplant melongena). These simple highly efficient great potential generating genome-edited plants crops.

Language: Английский

Citations

15
RNA virus-mediated gene editing for tomato trait breeding DOI Creative Commons
Mireia Uranga, Verónica Aragonés, Arcadio García

et al.

Horticulture Research, Journal Year: 2023, Volume and Issue: 11(1)

Published: Dec. 19, 2023

ABSTRACT Virus-induced genome editing (VIGE) leverages viral vectors to deliver CRISPR-Cas components into plants for robust and flexible trait engineering. We describe here a VIGE approach applying an RNA vector based on potato virus X (PVX) of tomato, mayor horticultural crop. Viral delivery single-guide Cas9-expressing lines resulted in efficient somatic with indel frequencies up 58%. By proof-of-concept PHYTOENE DESATURASE (PDS) plant regeneration from edited tissue, we recovered loss-of-function pds mutant progeny displaying albino phenotype. STAYGREEN 1 (SGR1), gene involved fruit color variation, generated sgr1 recolored red-brown fruits high chlorophyll levels. The obtained events were heritable, overall confirming the successful breeding color. Altogether, our offers great potential accelerated functional genomics tomato as well precision novel traits.

Language: Английский

Citations

14
Social and biological innovations are essential to deliver transformative forest biotechnologies DOI Open Access
Wout Boerjan, Steven H. Strauss

New Phytologist, Journal Year: 2024, Volume and Issue: 243(2), P. 526 - 536

Published: May 27, 2024

Summary Forests make immense contributions to societies in the form of ecological services and sustainable industrial products. However, they face major challenges their viability economic use due climate change growing biotic threats, for which recombinant DNA (rDNA) technology can sometimes provide solutions. But application rDNA technologies forest trees faces social biological obstacles that its societal acceptance a ‘wicked’ problem without straightforward We discuss nature these problems, innovations we consider essential progress. As case studies challenges, focus on modifications wood chemistry transformation efficiency. call regulations, dissolution method‐based market barriers, together could lead greater research investments, enable wide field studies, facilitate integration rDNA‐modified into conventional breeding programs. Without near‐term adoption such innovations, rDNA‐based solutions will be largely unavailable help forests adapt stresses from proliferation pests, nor available environmental benefits expanded related bioproducts as part an expanding bioeconomy.

Language: Английский

Citations

5
Viral delivery of an RNA-guided genome editor for transgene-free plant germline editing DOI Creative Commons
Trevor Weiss, Maris Kamalu, Honglue Shi

et al.

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

Published: July 19, 2024

Abstract Genome editing is transforming plant biology by enabling precise DNA modifications. However, delivery of systems into plants remains challenging, often requiring slow, genotype-specific methods such as tissue culture or transformation. Plant viruses, which naturally infect and spread to most tissues, present a promising system for reagents. But viruses have limited cargo capacities, restricting their ability carry large CRISPR-Cas systems. Here, we engineered tobacco rattle virus (TRV) the compact RNA-guided TnpB enzyme ISYmu1 its guide RNA. This innovation allowed transgene-free Arabidopsis thaliana in single step, with edits inherited subsequent generation. By overcoming traditional reagent barriers, this approach offers novel platform genome editing, can greatly accelerate biotechnology basic research.

Language: Английский

Citations

5
Heritable Tissue-Culture-Free Gene Editing in Nicotiana benthamiana through Viral Delivery of SpCas9 and sgRNA DOI Creative Commons
Tetsuya Yoshida, Masayuki Ishikawa, Seiichi Toki

et al.

Plant and Cell Physiology, Journal Year: 2024, Volume and Issue: 65(11), P. 1743 - 1750

Published: Aug. 31, 2024

Conventional plant gene editing requires laborious tissue-culture-mediated transformation, which restricts the range of applicable species. In this study, we developed a heritable and tissue-culture-free method in Nicotiana benthamiana using tobacco ringspot virus (TRSV) as vector for planta delivery Cas9 single-guide RNA (sgRNA) to shoot apical meristems. Agrobacterium-mediated inoculation TRSV induced systemic PHYTOENE DESATURASE. Transient downregulation silencing enhanced efficiency, resulting an order magnitude increase (0.8-13.2%) frequency transgenerational editing. While system had preference certain sgRNA sequences, co-inoculation carrying only rattle successfully introduced mutations with all five tested sgRNAs. Extensively gene-edited lateral shoots occasionally grew from plants inoculated vectors, ranged up 100%. This virus-mediated makes easy, requiring non-transgenic vector(s) obtain individuals.

Language: Английский

Citations

5
Advances and Prospects of Virus-Resistant Breeding in Tomatoes DOI Open Access

Zolfaghar Shahriari,

Xiaoxia Su, Kuanyu Zheng

et al.

International Journal of Molecular Sciences, Journal Year: 2023, Volume and Issue: 24(20), P. 15448 - 15448

Published: Oct. 22, 2023

Plant viruses are the main pathogens which cause significant quality and yield losses in tomato crops. The important that infect tomatoes worldwide belong to five genera: Begomovirus, Orthotospovirus, Tobamovirus, Potyvirus, Crinivirus. Tomato resistance genes against viruses, including Ty gene begomoviruses, Sw orthotospoviruses, Tm tobamoviruses, Pot 1 potyviruses, have been identified from wild germplasm introduced into cultivated cultivars via hybrid breeding. However, these mainly exhibit qualitative mediated by single genes, cannot protect virus mutations, recombination, mixed-infection, or emerging thus posing a great challenge antiviral Based on epidemic characteristics of we propose future studies breeding should focus rapidly, safely, efficiently creating broad-spectrum materials resistant multiple viruses. Accordingly, summarized analyzed advantages three strategies, marker-assisted selection (MAS)-based breeding, RNA interference (RNAi)-based transgenic CRISPR/Cas-based editing. Finally, highlighted challenges provided suggestions for improving using strategies.

Language: Английский

Citations

12
Engineering good viruses to improve crop performance DOI
Fabio Pasin, Mireia Uranga, R. Charudattan

et al.

Nature Reviews Bioengineering, Journal Year: 2024, Volume and Issue: 2(7), P. 532 - 534

Published: May 15, 2024

Language: Английский

Citations

4