Tripathi Sushil K, Gupta Kailash C, Kumar Pradeep
Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi-110 007, India.
Mol Biosyst. 2013 Sep;9(9):2322-30. doi: 10.1039/c3mb70150f.
A series of electrostatically crosslinked nanoparticles, N-(2-hydroxyethyl)-polyethylenimine-PEG600 (HePP), was prepared by allowing N-(2-hydroxyethyl)-polyethylenimine (HeP) to interact with polyethyleneglycol (600) dicarboxylic acid (HOOC-PEG600-COOH, PEG600dc), they were then evaluated for their capability to transfect cells in vitro and in vivo. DLS studies revealed the size of the HePP nanoparticles in the range 106-170 nm, which efficiently condensed nucleic acids and provided sufficient protection against nuclease degradation. HePP-pDNA complexes exhibited a considerably higher transfection efficiency and cell viability in various mammalian cell lines, with HePP-3-pDNA displaying the highest gene expression, which outperformed HeP and the commercially available transfection reagent, Lipofectamine™. Also, HePP-3 mediated sequential delivery of GFP specific siRNA resulted in ∼76% suppression of the target gene. Intravenous administration of HePP-3-pDNA complex to mice, followed by monitoring of the reporter gene analysis post 7d, revealed the highest gene expression occurred in the spleen. Together, these results advocate the potential of HePP nanoparticles as efficient vectors for gene delivery in vitro and in vivo.
通过使N-(2-羟乙基)聚乙烯亚胺(HeP)与聚乙二醇(600)二羧酸(HOOC-PEG600-COOH, PEG600dc)相互作用，制备了一系列静电交联纳米颗粒N-(2-羟乙基)聚乙烯亚胺-PEG600(HePP)，然后评估它们在体外和体内转染细胞的能力。动态光散射研究表明，HePP纳米颗粒的尺寸在106-170nm范围内，其能有效凝聚核酸并提供足够的保护以抵抗核酸酶降解。HePP-pDNA复合物在各种哺乳动物细胞系中表现出相当高的转染效率和细胞活力，其中HePP-3-pDNA表现出最高的基因表达，其性能优于HeP和市售转染试剂Lipofectamine™。此外，HePP-3介导的绿色荧光蛋白特异性小干扰RNA的顺序递送导致靶基因约76%的抑制。给小鼠静脉注射HePP-3-pDNA复合物，然后在7天后监测报告基因分析，结果显示脾脏中出现最高的基因表达。总之，这些结果表明HePP纳米颗粒作为体外和体内基因递送的有效载体具有潜力。
