Aug 2026· Frontiers in Plant Science· 0 citations· 164 references
TL;DR
This review provides a comprehensive synthesis of a recent advances in CRISPR–Cas technologies and their strategic applications in crop genetics and hybrid breeding, and showcases how these technologies accelerate hybrid breeding by engineering male sterility systems, fixing heterosis, and generating high-throughput mutant libraries for trait discovery.
Abstract
Developing climate-smart crops with enhanced crop productivity, nutritional quality, resistance to biological and environmental stressors is vital for global food security. While hybrid breeding forms the cornerstone of modern crop improvement, conventional breeding approaches are limited by genetic barriers and prolonged breeding cycles. CRISPR–Cas based genome editing has revolutionized plant biology by allowing precise, efficient, and multiplex genetic modifications. This review provides a comprehensive synthesis of a recent advances in CRISPR–Cas technologies and their strategic applications in crop genetics and hybrid breeding. We summarize major genome-editing strategies, including gene knock-out, base editing (BE), knock-in, gene replacement, epigenome editing, and transcriptional regulation. Furthermore, we contrast stable, transient, and DNA-free delivery systems, highlighting ribonucleoprotein (RNP)-mediated delivery for minimizing off-target effects and avoiding transgene integration. We showcase how these technologies accelerate hybrid breeding by engineering male sterility systems, fixing heterosis, and generating high-throughput mutant libraries for trait discovery. Finally, we synthesize major bottlenecks in tissue culture-independent transformation and delivery systems, while outlining how emerging paradigms like
de novo
domestication and synthetic biology will shape the future of climate-resilient agriculture.
The application of genome editing, CRISPR/Cas9 has revolutionized plant breeding by enabling precise, efficient, and targeted modification of native genes, significantly accelerating the development of improved agronomic traits of crops. Therefore, CRISPR/Cas9 technology currently the most extensively used genome editing technique worldwide because of its simple design, cost-effectiveness, high efficiency, good reproducibility, high engineering feasibility, ability to create gene knockout, RNA editing, and quick cycle. It is used to knock in or knock out genes of interest and for generating models for genetic studies. The main components of the CRISPR/Cas9 system are an RNA-guided Cas9 endonuclease and a single-guide RNA (sgRNA). The workflow of CRISPR/Cas9 gene editing comprises selecting target sites, designing and synthesizing sgRNA, introducing transformation constructs or ribonucleoprotein (RNP) in plant cells, followed by transformation and identification of edited lines. This approach bypasses the formal regulations on GMOs, thus encouraging the widespread adoption RNA-guided gene editing in agricultural sciences and biotechnology. The system is now being utilized in the biofortification of cereal crops such as rice, wheat, barley, and maize, including vegetable crops such as potato and tomato. The world's first genome-edited rice varieties are DRR Dhan 100 (Kamala) and Pusa DST Rice 1 developed by the Indian Council of Agricultural Research (ICAR), New Delhi, India in 2025 with the objective of bringing about revolutionary changes in terms of higher production, climate adaptability, and water conservation. The CRISPR/Cas9-based crop genome editing has been utilized in imparting/producing qualitative enhancement in aroma, shelf life, sweetness, and quantitative improvement in starch, protein, gamma-aminobutyric acid (GABA), oleic acid, anthocyanin, phytic acid, gluten, and steroidal glycoalkaloid contents. Some varieties have even been modified to become disease and stress-resistant. Therefore, CRISPR/Cas9 is aiding in developing climate-ready crops and improving crop quality parameters such as appearance, palatability, nutritional components, and other preferred traits. Gene editing tools are used to generate changes to the native genetic material. Unlike GMOs, which introduce novel configurations of genetic materials typically derived from other organisms, gene editing methods modify existing genetic material in ways that can yield beneficial outcomes.
Ravindra B. Malabadi, Raju K. Chalannavar· World Journal of Advanced Re...· 0 citations
CRISPR/Cas-based genome editing has emerged as a powerful and precise tool for crop improvement, enabling targeted modification of genes associated with agriculturally important traits. In horticultural crops, CRISPR technologies have accelerated the improvement of disease resistance, abiotic stress tolerance, yield, nutritional quality, shelf life, flowering behavior, and ornamental characteristics. Among available genome-editing platforms, CRISPR/Cas9 is the most widely utilized because of its simplicity, efficiency, and versatility. The technology enables precise genome modification through targeted DNA cleavage followed by endogenous repair mechanisms, facilitating gene knockout, insertion, or sequence alteration. Recent advances in genome editing have significantly expanded its applications in vegetable, fruit, and ornamental crops. Successful modifications targeting genes associated with stress tolerance, fruit ripening, pigment biosynthesis, flowering regulation, and pathogen resistance demonstrate the enormous potential of CRISPR-mediated breeding for horticultural improvement. However, several challenges, including low transformation efficiency, genotype-dependent regeneration, prolonged juvenile phases, polyploidy, and regulatory concerns, continue to limit its broader application in many horticultural species. This review summarizes recent progress in CRISPR/Cas-mediated genome editing in horticultural crops, including strategies for guide RNA design, transformation, regeneration, development of transgene-free plants, and regulatory considerations. Furthermore, emerging advances such as precision editing technologies and improved delivery systems are discussed as promising approaches for enhancing editing efficiency and expanding future applications. Overall, CRISPR/Cas technologies hold substantial potential for accelerating the development of climate-resilient, high-quality, and nutritionally improved horticultural crops.
Prerna Srivastava, D. Singh, Rima Kumari et al.· Discover Plants· 0 citations
This review critically synthesizes recent advances in CRISPR applications for major wheat fungal diseases, including powdery mildew, rusts, Fusarium head blight, and wheat blast, and highlights future opportunities for integrating genome editing with modern breeding to accelerate the development of climate-resilient, disease-resistant wheat cultivars for sustainable agriculture.
M. S. Samoo· Bulletin of the National Res...· 1 citation
Overall, the evidence suggests that CRISPR Cas-9 has the potential to accelerate plant breeding, improve food security, and support the development of climate resilient crops while reducing the time and uncertainty associated with traditional breeding methods.
Victoria Imafidor· World Journal of Advanced En...· 0 citations
The future of crop improvement using GEd technologies lies in the harmonisation or alignment of global policies and regulations to support the trade of agricultural produce and ensure that growers and consumers can benefit from GEd technology.
Michael G. K. Jones· Sugar Industry international· 0 citations