International Journal of Horticulture, 2025, Vol.15, No.3, 113-122 http://hortherbpublisher.com/index.php/ijh 115 are more acceptable to consumers and regulatory bodies. The image illustrates the target selection process in a CRISPR tool, using online tools such as CRISPR-direct and CRISPR-P to design a suitable target site. This process ensures efficient and specific identification of editing targets in the potato genome. Sequence selection of target genes and design of sgRNAs are critical steps that provide the basis for subsequent gene editing. In the construction of gene editing vectors, researchers integrate sgRNA and Cas9 nuclease into the expression vector to achieve efficient editing efficiency. These vectors can be introduced into potato cells using a variety of transformation methods, and panel D illustrates the main transformation methods including protoplast transfection, Agrobacterium-mediated transformation, gene gunning, and viral vector infection. These methods are suitable for specific research needs due to their different advantages and disadvantages. At the heart of gene editing is the synergistic effect of Cas9 nuclease with sgRNA, which allows researchers to knock out, repair, or replace target genes through non-homologous end joining (NHEJ) or homologous recombination (HDR). This precise editing capability makes it possible to develop more resistant, higher-yielding, or higher-quality potato varieties. For example, potato resistance to late blight can be improved by knocking out genes associated with disease susceptibility (Figure 1) (Tussipkan and Manabayeva, 2021). CRISPR-Cas9 has been used to develop potato varieties with resistance to the Colorado potato beetle and late blight, as well as to reduce acrylamide content and modify starch composition. The transient expression of genome editing components in potato protoplasts has also been reported to generate edited plants without the integration of foreign DNA, which is a significant advancement from both scientific and regulatory perspectives (Li et al., 2024). Figure 1 Schematic representation of plant genome editing with CRISPR/Cas (Adopted from Tussipkan and Manabayeva, 2021)
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