Tomato (Solanum lycopersicum) is a premier global crop, celebrated not only for its economic importance but also as a fundamental “well-being food” that provides essential vitamins, minerals, and potent antioxidants. This review highlights the transformative impact of CRISPR/Cas9 and next-generation precision tools, such as base and prime editing, on modern tomato breeding. Beyond functional genomics, we focus on the strategic engineering of tomato to maximize productivity through fruit size optimization and the biofortification of high-value metabolites (e.g., GABA, lycopene, and vitamins). Furthermore, we examine recent advances in developing multi-stress resilience against both biotic and abiotic challenges. A significant emphasis is placed on re-designing plant architecture and developmental traits to tailor tomato varieties for future agricultural systems, particularly indoor and vertical farming. By advanced delivery methods, CRISPR technologies offer unprecedented potential to accelerate the development of “smart” tomato, ensuring sustainable agriculture and global food security in a changing climate.

Cadmium (Cd) contamination in rice poses significant health risks to consumers. This study aimed to reduce Cd accumulation in the elite Vietnamese rice variety TBR225 (TBR225) by editing the Natural Resistance-Associated Macrophage Protein 5 (OsNRAMP5) gene using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR- associated 9 (Cas9) technology. We successfully generated OsNRAMP5 knockout mutants through Agrobacterium-mediated transformation. Sequencing analysis revealed various mutation types, including deletions, insertions, and substitutions, all resulting in premature stop codons or frameshift mutations. Three homozygous, transgene-free mutant lines were selected for further analysis. These lines exhibited significantly reduced Cd accumulation in roots (78.4-84.5%), shoots (72.3-83.8%), and grains (50.5-66.0%) compared to wild-type plants when exposed to different Cd concentrations. Importantly, the OsNRAMP5 mutations did not adversely affect major agronomic traits, including growth time, plant height, tiller number, grain yield, or amylose content. Additionally, the accumulation of essential micronutrients such as Fe and Zn remained unaffected in the mutant lines. Our results demonstrate the potential of CRISPR/Cas9-mediated OsNRAMP5 editing as an effective strategy for developing low-Cd accumulating rice varieties without compromising agronomic performance or nutritional quality. This approach could significantly contribute to enhancing food safety in regions facing Cd contamination challenges.

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Targeted genome editing using CRISPR/Cas nucleases has become the standard approach for creating mutant plants. Significant progress has been made to enhance the editing efficiencies through optimizing CRISPR/Cas expression, including applying heat stress. In this study, we used heat stress to enhance the Potato virus X (PVX)-mediated CRISPR/Cas9 mutagenesis in Nicotiana benthamiana. We show that heat stress at 4-5 days after PVX inoculation effectively increases the mutagenesis efficiency of Cas9 nuclease. We observed up to a 5-8% increase in mutation efficiency depending on the sgRNA construct when heat stress is applied to the pPVX-Cas9::sgRNA infiltrated samples. Furthermore, analysis of the effect of the heat stress on the pattern of mutation types in the target gene regions showed no obvious changes in CRISPR/Cas9 induced mutagenesis pattern between heat stress treated and no heat stress treated samples. Overall, our experiments demonstrate that heat stress treatment at the optimal time after viral inoculation is most effective in increasing the PVX-mediated CRISPR/Cas9 editing efficiency in plants.

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Crop improvement is essential to attaining world food security and enhancing nutrition for human beings. Both conventional breeding and modern molecular breeding have contributed to increased crop production and quality. However, the time and resources for breeding practices have been limited. It takes a long time to bring a novel improved crop to the market, and the genetic sources from wild species cannot be always available for crops of our interests. Genome editing technology implemented molecular breeding can overcome those limitations of time and resource by facilitating the specific editing of plant genomes. However, there is a long-lasting argument about the safety of genetically modified organisms (GMOs). In this review, we briefly summarize the principle of genome editing tools, focusing on the CRISPR/Cas9 system and the application of these tools to plants in the service of crop engineering.

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