Cannabis sativa (Cannabis) is a multipurpose plant species consisting of specific lineages that for centuries has either been artificially selected for the production of fiber or the psychoactive drug Δ⁹-tetrahydrocannabinol (THC). With the recent lifting of previous legal restrictions on consuming Cannabis, there has been a resurgence of interest in understanding and manipulating Cannabis genetics to enhance its compositions. Yet, recently developed approaches are not amenable to high-throughput gene stacking to study multi-genic traits. Here, we demonstrate an efficient nanoparticle-based transient gene transformation protocol where multiple gene plasmids can be expressed simultaneously in intact Cannabis leaf cells in a very short time (5 days). Constructs encoding two soybean transcription factors were co-grafted onto poly-ethylenimine cationic polymer-modified silicon dioxide-coated gold nanoparticles (PEI-Au@SiO₂). Infiltration of the DNA-PEI-Au@SiO₂ into Cannabis leaf tissues resulted in the transcription of both soybean genes and the localization of fluorescent-tagged transcription factor proteins in the nuclei of Cannabis leaf cells including the trichomes, which are the cell types that biosynthesize valuable cannabinoid and terpene metabolites. Our study exemplifies a rapid transient gene transformation approach that will be useful to study the effects of gene stacking in Cannabis.

大麻(Cannabis) 是一种多用途植物物种，由特定谱系组成，几个世纪以来，这些谱系被人工选择用于生产纤维或精神活性药物 Δ ⁹-四氢大麻酚 (THC)。随着最近对消费大麻的先前法律限制的解除，人们对理解和操纵大麻遗传学以增强其成分的兴趣重新燃起。然而，最近开发的方法不适用于高通量基因堆叠来研究多基因性状。在这里，我们展示了一种有效的基于纳米颗粒的瞬时基因转化方案，其中多个基因质粒可以在很短的时间内（5 天）在完整的大麻叶细胞中同时表达。将编码两种大豆转录因子的构建体共接枝到聚乙烯亚胺阳离子聚合物改性的二氧化硅包覆的金纳米粒子 (PEI-Au@SiO ₂ ) 上。DNA-PEI-Au@SiO _{2 的}渗透进入大麻叶组织导致大豆基因的转录和荧光标记的转录因子蛋白在包括毛状体在内的大麻叶细胞核中的定位，毛状体是生物合成有价值的大麻素和萜烯代谢物的细胞类型。我们的研究举例说明了一种快速瞬时基因转化方法，这将有助于研究大麻中基因堆积的影响。