A novel protocol for protoplast isolation, transfection, and culture in Cannabis sativa L

Abstract Background: Protoplasts are a valuable tool for studying gene expression and applying genome editing techniques. Given the high medicinal and industrial potential of Cannabis sativa L., developing an efficient protoplast-to-plant regeneration protocol is highly desirable. Due to its recalcitrant nature, a complete plant regeneration from cannabis protoplasts has not yet been achieved. Results: This study details a robust protocol for cannabis protoplast isolation, purification, transient transfection, and culture, additionally reporting somatic embryo-like structures derived from protoplast-derived callus. We demonstrated