介孔硅纳米粒作为植物细胞转基因载体的研究

Genetic Transformation of Plant Cells Mediated by Mesoporous Silica Nanoparticles

随着纳米生物技术的快速发展,以纳米材料作为基因载体在植物细胞转导研究中取得了初步进展。本研究制备了两种尺寸的介孔硅纳米颗粒,氨基修饰后对其进行表征,并负载含smGFP基因的质粒DNA对拟南芥原生质体进行细胞转导。结果表明,直径约20 nm的氨基化介孔硅纳米颗粒(Am-MSN-20)呈类球形,花形结构的氨基化介孔硅纳米颗粒(Am-MSN-50)直径约50 nm,两者均带正电荷。Am-MSN-50结合DNA的能力高于Am-MSN-20,两种纳米载体都表现出了良好的稳定性,能够保护负载的pDNA不被细胞核酸酶降解,并且对原生质体没有毒害作用。与Am-MSN-20相比较,Am-MSN-50具备更高的转导效率。本研究表明,氨基修饰的MSNs有望成为一种安全高效的新型植物基因载体。

With the rapid development of nanobiotechnology,nanoparticles have been applied as vectors for gene transformation in plant cells.In this study,mesoporous silica nanoparticles(MSNs)with different particle sizes were synthesized and then amine-functionalized.The transformation efficiencies of amine-functionalized MSNs(Am-MSNs)for delivering plasmid DNA encoding green fluorescent protein(smGFP)were investigated in the protoplasts of Arabidopsis thaliana.As results,the Am-MSN-20 exhibited monodispersed spheroids with average particle size of 20 nm,while Am-MSN-50 showed a flower-like structure and had an average particle size of 50 nm.Both amine-functionalized MSNs were positively charged.Am-MSN-50 had higher binding capacity of pDNA than that of Am-MSN-20.The Am-MSNs/pDNA complex showed good stability,which suggested that both Am-MSNs protected bounded pDNA efficiently against degradation by cellular nucleases.The MSNs also exhibited no cytotoxic effects on the protoplasts of A.thaliana.The Am-MSN-50 enabled much higher transformation efficiency of the pDNA encoding smGFP compared to that of Am-MSN-20.The results suggest that Am-MSNs can be employed as promising carriers for safe and effective gene delivery.