CRISPR/Cas-based precision gene replacement in plants via microhomology-mediated end joining DOI Open Access
Tien Van Vu, Gah‐Hyun Lim,

Seung Hee Choi

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Aug. 27, 2022

Abstract Precise gene or allele replacement is a desirable technology, but implementing it in plants remains challenging. CRISPR-Cas-based approaches, such as targeting (GT) and prime editing (PE), have opened up new possibilities for precise plants. However, their size efficiency still need improvement. Recently, strategies using canonical nonhomologous end-joining (cNHEJ) microhomology-mediated end joining (MMEJ) been considered promising alternatives yeast mammals. these approaches not extensively explored applied to Here, we proposed tested tool, termed PREMJ ( p recision re placement via m icrohomology-mediated joining), precision The strategy employing 20-bp microhomology MMEJ donors (∼100 bp lengths) inhibitor, NU7441, produced high targeted efficiencies, 1.60 ± 0.14, 4.47 1.98, 8.98 4.73 % protoplasts of tomato, lettuce, cabbage, respectively. Our data also revealed the critical impacts length NU7441 concentration on PREMJ-based Although obtaining edited challenging due inefficient protoplast regenerations Agrobacterium -mediated delivery, may significantly contribute competent complex delivery plant regeneration. Key Message We designed method, PREMJ, CRISPR-Cas-mediated double-stranded break (DSB) formation repairing DSBs with donor template carrying desired base changes microhomologies flanking DSB ends. showed feasibility tomatoes, albeit its efficacy transformation requires further optimization.

Language: Английский

Engineering source-sink relations by prime editing confers heat-stress resilience in tomato and rice DOI

H. F. Lou,

Shujia Li,

Zihang Shi

et al.

Cell, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 1, 2024

Language: Английский

Citations

12
Genetic Improvement of rice Grain size Using the CRISPR/Cas9 System DOI Creative Commons
Tao Zhang, Zhengwei Wang, Qiaoquan Liu

et al.

Rice, Journal Year: 2025, Volume and Issue: 18(1)

Published: Jan. 27, 2025

Rice grain size influences both yield and quality, making it a significant target for rice genetic improvement. In recent years, numerous genes related to with differential effects have been cloned. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing system is convenient tool modifying genes. use of the CRISPR/Cas9 improvement size-related worth exploring. This paper summarizes known modification discusses potential applications improving size.

Language: Английский

Citations

1
CRISPR–Cas applications in agriculture and plant research DOI
Aytug Tuncel, Changtian Pan,

Joshua S. Clem

et al.

Nature Reviews Molecular Cell Biology, Journal Year: 2025, Volume and Issue: unknown

Published: March 7, 2025

Language: Английский

Citations

1
Perfecting prime editing: achieving precise edits in dicots DOI
Niaz Ahmad, Muhammad Jawad Akbar Awan, Imran Amin

et al.

Trends in Plant Science, Journal Year: 2025, Volume and Issue: 30(3), P. 232 - 234

Published: Jan. 2, 2025

Language: Английский

Citations

0
CRISPR/Cas-Mediated Genetic Improvements in Vegetable Crops DOI

Noman Nazik,

Anowarul Amin,

Zujun Yin

et al.

Published: Jan. 1, 2025

Language: Английский

Citations

0
CRISPR-mediated genome editing of wheat for enhancing disease resistance DOI Creative Commons
Joshua Waites, V. Mohan Murali Achary, Easter D. Syombua

et al.

Frontiers in Genome Editing, Journal Year: 2025, Volume and Issue: 7

Published: Feb. 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.

Language: Английский

Citations

0
Applying conventional and cell-type-specific CRISPR/Cas9 genome editing in legume plants DOI Creative Commons
Jin‐Peng Gao, Yangyang Su, Suyu Jiang

et al.

aBIOTECH, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 16, 2024

Abstract The advent of genome editing technologies, particularly CRISPR/Cas9, has significantly advanced the generation legume mutants for reverse genetic studies and understanding mechanics rhizobial symbiosis. legume–rhizobia symbiosis is crucial sustainable agriculture, enhancing nitrogen fixation improving soil fertility. Numerous genes with a symbiosis-specific expression have been identified, sometimes exclusively expressed in cells forming infection threads or nitrogen-fixing nodule cells. Typically, mutations these do not affect plant growth. However, some instances, germline homozygous can be lethal result complex pleiotropic phenotypes that are challenging to interpret. To address this issue, rhizobia-inducible cell-type-specific CRISPR/Cas9 strategy was developed knock-out specific transgenic root tissues. In review, we discuss recent advancements editing, highlighting CRISPR system its applications symbiotic beyond.

Language: Английский

Citations

1
Enhancing CRISPR-Cas-based gene targeting in tomato using a dominant-negative ku80 DOI Creative Commons
Tien Van Vu,

Ngan Thi Nguyen,

Jihae Kim

et al.

Horticulture Research, Journal Year: 2024, Volume and Issue: 12(2)

Published: Oct. 23, 2024

Abstract The CRISPR-Cas-based gene targeting (GT) method has enabled precise modifications of genomic DNA ranging from single base to several kilobase scales through homologous recombination (HR). In plant somatic cells, canonical non-homologous end-joining (cNHEJ) is the predominant mechanism for repairing double-stranded breaks (DSBs), thus limiting HR-mediated GT. this study, we implemented an approach shift repair pathway preference toward HR by using a dominant-negative ku80 mutant protein (KUDN) disrupt initiation cNHEJ. employment KUDN conferred 1.71- 3.55-fold improvement in GT efficiency at callus stage. When screened transformants, there was more remarkable increase efficiency, 1.62- 9.84-fold, two specific tomato loci, SlHKT1;2 and SlEPSPS1. With practical levels enhanced KUDN-based tool successfully facilitated 9-bp addition additional locus, SlCAB13. These findings provide another promising efficient breeding.

Language: Английский

Citations

0
Applications and Prospects of CRISPR/Cas9 Technology in the Breeding of Major Tropical Crops DOI Creative Commons
Lixia Zhou, Xianhai Zeng, Yaodong Yang

et al.

Plants, Journal Year: 2024, Volume and Issue: 13(23), P. 3388 - 3388

Published: Dec. 2, 2024

China is a major producer of tropical crops globally, boasting rich varieties and diverse functions. Tropical account for two-thirds the plant species in this country. Many their products, such as oil palm, rubber, banana, sugarcane, cassava, papaya are well known to people. Most these products irreplaceable possess special They not only supply important raw materials people's daily life industrial agricultural production but also contribute economic growth subtropical regions China. However, modern molecular breeding severely hampered by biological characteristics genetic complexity. Issues polyploidy, heterozygosity, vegetative propagation, long juvenile periods, large sizes result time consuming, low efficiency, slow progress conventional crops. The development genome-editing technologies has brought new way breeding. As an emerging gene-editing technology, CRISPR-Cas9 system been widely used plants, adopted its higher targeting versatility, ease usage. This approach applied papaya. review summarized delivery patterns, mutation detection, application crop breeding, discussed existing problems, addressed prospects future applications field, providing references relevant studies.

Language: Английский

Citations

0
CRISPR/Cas-based precision gene replacement in plants via microhomology-mediated end joining DOI Open Access
Tien Van Vu, Gah‐Hyun Lim,

Seung Hee Choi

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Aug. 27, 2022

Abstract Precise gene or allele replacement is a desirable technology, but implementing it in plants remains challenging. CRISPR-Cas-based approaches, such as targeting (GT) and prime editing (PE), have opened up new possibilities for precise plants. However, their size efficiency still need improvement. Recently, strategies using canonical nonhomologous end-joining (cNHEJ) microhomology-mediated end joining (MMEJ) been considered promising alternatives yeast mammals. these approaches not extensively explored applied to Here, we proposed tested tool, termed PREMJ ( p recision re placement via m icrohomology-mediated joining), precision The strategy employing 20-bp microhomology MMEJ donors (∼100 bp lengths) inhibitor, NU7441, produced high targeted efficiencies, 1.60 ± 0.14, 4.47 1.98, 8.98 4.73 % protoplasts of tomato, lettuce, cabbage, respectively. Our data also revealed the critical impacts length NU7441 concentration on PREMJ-based Although obtaining edited challenging due inefficient protoplast regenerations Agrobacterium -mediated delivery, may significantly contribute competent complex delivery plant regeneration. Key Message We designed method, PREMJ, CRISPR-Cas-mediated double-stranded break (DSB) formation repairing DSBs with donor template carrying desired base changes microhomologies flanking DSB ends. showed feasibility tomatoes, albeit its efficacy transformation requires further optimization.

Language: Английский

Citations

1