Summary: Hemp (CannabissativaL.) is an annual and typically dioecious crop. Due to the therapeutic potential for human diseases, phytocannabinoids as a medical therapy is getting more attention recently. Several candidate genes involved in cannabinoid biosynthesis have been elucidated using omics analysis. However, the gene function was not fully validated due to few reports of stable transformation for Cannabis tissues. In this study, we firstly report the successful generation of gene‐edited plants using anAgrobacterium‐mediated transformation method inC.sativa. DMG278 achieved the highest shoot induction rate, which was selected as the model strain for transformation. By overexpressing the cannabis developmental regulator chimera in the embryo hypocotyls of immature grains, the shoot regeneration efficiency was substantially increased. We used CRISPR/Cas9 technology to edit the phytoene desaturase gene and finally generated four edited cannabis seedlings with albino phenotype. Moreover, we propagated the transgenic plants and validated the stable integration of T‐DNA in cannabis genome.

Cannabis (Cannabis sativa L.) is a dioecious plant cultivated worldwide for thousands of years. Besides the narcotic and therapeutic effects, Cannabis can be used as raw materials in multiple fields, including bioenergy, textiles, food, and ecological restoration. It is also an efficient bioremediation agent for contaminated soil, as well as greenhouse gas absorption. With the expansion of the market, there has been an increased demand to develop Cannabis cultivars with enhanced traits. As a major science breakthrough, the advent of the CRISPR/Cas system will revolutionize the basic and applied research in Cannabis. This article provides an overview of the recent advances in the optimization of a transformation system and in the gene editing of Cannabis. To achieve the full potential of this environmentally friendly and sustainable crop, we highlight future directions of genetic modification as well as several bottlenecks to overcome.