Grafting has been utilized to demonstrate the long-distance transportation of molecules, including hormones, proteins, and RNA. The fusion of CRISPR/Cas9 sequences with tRNA-like sequences (TLS) enables the generation of transgenic plants capable of producing mobile CRISPR/Cas9 RNA. This technology holds promise for enhancing the efficiency of generating diverse gene-edited plants via grafting. Our study aimed to identify optimal grafting conditions in tomatoes by adjusting both the plant's developmental stage and the diameter of the grafting clip. Grafting experiments were conducted on different varieties of tomatoes, as well as on other species such as tobacco and tomatoes. Grafting clips with diameters of 2-3 mm were employed, with tomatoes selected between 30 and 60 days after sowing. Optimal grafting conditions were achieved when tomatoes were grafted between 45 and 50 days post-sowing, prior to flowering, using 2.5 mm clips. Gene editing through grafting offers the advantage of producing transgene-free offspring in a single generation. We aim to employ these optimized grafting conditions for the generation of gene-edited tomatoes. Specifically, we targeted the tomato DR genes, which is associated with enhanced vitamin D content. Micro-Tom variety with high transformation efficiency was used in the experiment. Consequently, our strategy targets the production of tomato cultivars that enhance Vitamin D levels through a combination of plant crossbreeding and grafting techniques.