CRISPR/Cas9 gene editing system is recently developed robust genome editing technology for accelerating plant breeding. Various modifications of this editing system have been established for adaptability in plant varieties as well as for its improved efficiency and portability. This review provides an in-depth look at the various strategies for synthesizing gRNAs for efficient delivery in plant cells, including chemical synthesis and in vitro transcription. It also covers traditional analytical tools and emerging developments in detection methods to analyze CRISPR/Cas9 mediated mutation in plant breeding. Additionally, the review outlines the various analytical tools which are used to detect and analyze CRISPR/Cas9 mediated mutations, such as next-generation sequencing, restriction enzyme analysis, and southern blotting. Finally, the review discusses emerging detection methods, including digital PCR and qPCR. Hence, CRISPR/Cas9 has great potential for transforming agriculture and opening avenues for new advancements in the system for gene editing in plants.

CRISPR/Cas‐based genome editing has emerged as a transformative tool for precise genetic improvement of cereal crops. Recent advances in CRISPR technologies, including Cas9, Cas12, Cas13, base editing, and prime editing, have enabled targeted modification of genes and regulatory elements controlling yield, stress tolerance, and grain nutritional quality in major cereals such as rice, wheat, maize, and barley. This review summarizes current progress in CRISPR‐mediated genome editing systems, delivery strategies, and representative applications in cereal crop improvement. Emphasis is placed on how genome editing reprograms enzymatic activities and biological pathways underlying complex agronomic traits rather than acting through single‐gene effects. The review also discusses challenges related to trait complexity, regulatory considerations, and prospects for translating genome‐edited cereal crops from laboratory research to field‐level application. Collectively, this review highlights the potential of CRISPR/Cas genome editing as a powerful approach for developing high‐yielding, resilient, and nutritionally improved cereal crops.

Clustered regularly interspaced short palindromic repeats/CRISPR‐associated protein 9 (CRISPR/Cas9) stands as a major technique in genetic engineering, offering innumerable applications for enhancing agricultural and horticultural crops. It contributes significantly to improving quality traits, managing biotic and abiotic stresses, and extending shelf life. Several genes, such as Cry genes or protease inhibitors and trypsin inhibitors, have been utilized to confer resistance against insect pests and diseases. This technique finds extensive application in medicine, agricultural, and horticultural crops. CRISPR/Cas9 has been successfully implemented in tomato, petunia, grapes, apple, and citrus, yielding promising results. For instance, in tomato, CRISPR/Cas9 has been employed with the Agrobacterium gene transfer method to provide resistance against powdery mildew disease. This review presents a comprehensive overview of CRISPR/Cas9 applications and achievements in agricultural and horticultural crops. Overall, this review highlights the potential applications of CRISPR technology in plant improvement, with limitations and challenges including off‐target mutations, size of the CRISPR/Cas9 system, delivery, and bio‐safety regulations to improve a variety of crop traits, including yield, quality, stress tolerance, disease, and herbicide resistance. It can also be used to speed up crop improvement compared to conventional breeding.