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从植物到植物:植物源生物聚合物作为将DNA直接递送至植物细胞的可持续且安全的纳米载体

From Plants to Plants: Plant-Derived Biological Polymers as Sustainable and Safe Nanocarriers for Direct Delivery of DNA to Plant Cells.

作者信息

Vinzant Kari, Rashid Mohammad, Clouse Delaney E, Ghosh Pratyusha, Quadir Mohiuddin, Davis Virginia A, Khodakovskaya Mariya V

机构信息

University of Arkansas at Little Rock, Little Rock, Arkansas 72204, United States.

Auburn University, Auburn, Alabama 36849, United States.

出版信息

Nano Lett. 2025 Apr 9;25(14):5572-5581. doi: 10.1021/acs.nanolett.4c05489. Epub 2025 Mar 26.

Abstract

Sustainable plant-derived biopolymers cellulose nanocrystals (CNC) and Zein protein were used to deliver plasmid DNA with a reporter GFP gene (pDNA) to plant cells. CNC and Zein were modified with the cationic agent 2,3-epoxypropyltrimethylammonium chloride (EPTMAC) to electrostatically bind the biopolymers to negatively charged pDNA. Established pDNA-CNC and pDNA-Zeins conjugates were delivered to tobacco cells by leaf injection and vacuum infiltration of tobacco leaves and seedlings. Both methods effectively provided transient GFP expression in exposed plant cells that was visualized by confocal microscopy and confirmed by qRT-PCR (GFP gene expression) and Western blot (GFP protein expression). Our findings support the idea that nanopolymers derived from agricultural waste residues can successfully be used to advance plant transformation and gene editing. Delivering genetic material using biocompatible, plant-based nanopolymers in large-scale vacuum infiltration of plant tissues reduces existing limitations of plant transformation and increases the speed of the transformation process.

摘要

可持续的植物源生物聚合物纤维素纳米晶体(CNC)和玉米醇溶蛋白被用于将携带报告基因绿色荧光蛋白基因(pDNA)的质粒DNA递送至植物细胞。用阳离子试剂2,3-环氧丙基三甲基氯化铵(EPTMAC)对CNC和玉米醇溶蛋白进行修饰,以使生物聚合物与带负电荷的pDNA静电结合。通过叶片注射以及对烟草叶片和幼苗进行真空渗透,将已制备好的pDNA-CNC和pDNA-玉米醇溶蛋白缀合物递送至烟草细胞。两种方法均有效地在暴露的植物细胞中实现了绿色荧光蛋白的瞬时表达,这通过共聚焦显微镜观察得以可视化,并通过qRT-PCR(绿色荧光蛋白基因表达)和蛋白质免疫印迹法(绿色荧光蛋白表达)得到证实。我们的研究结果支持这样一种观点,即源自农业废弃物的纳米聚合物能够成功用于推进植物转化和基因编辑。在植物组织的大规模真空渗透中使用生物相容性的、基于植物的纳米聚合物递送遗传物质,减少了植物转化的现有局限性,并提高了转化过程的速度。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cc58/11987027/b9d3305be29d/nl4c05489_0001.jpg

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