High performance mRNA transfection through carbonate apatite-cationic liposome conjugates.
High performance mRNA transfection through carbonate apatite-cationic liposome conjugates.
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DOI：
10.1016/j.biomaterials.2009.02.050
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发表时间：
2009-08
期刊：
影响因子：
14
通讯作者：
F. Zohra;E. Chowdhury;T. Akaike
中科院分区：
文献类型：
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作者：
F. Zohra;E. Chowdhury;T. Akaike
mRNA代替DNA为基因治疗和基因疫苗接种提供了一种新的、有吸引力的方法。mRNA的传递可以绕过核定位步骤，通过转录直接在细胞质中表达蛋白质。目前的mRNA递送技术主要基于阳离子脂质体，转染活性低。我们之前报道过，将无机碳酸盐磷灰石纳米颗粒应用于阳离子脂质体DOTAP {N-[1-（2,3-二油氧基）丙基]-N，N，N-三甲基氯化铵}上，在有丝分裂细胞和非有丝分裂细胞中都能获得高转染荧光素酶mRNA的能力。在本文中，我们扩展了之前的工作，特别是在两个重要部分进行了详细的研究，即评估复合物的图像和分析基因传递的步骤，以检测增强转染效力的决定因素。透射电镜（TEM）观察清楚地表明，无机碳酸盐磷灰石颗粒存在于mrna -脂质体复合物上，并展示了新型杂化载体材料的结构。与DOTAP相比，由于吸收的无机纳米颗粒的重力明显增加，细胞接触和混合颗粒相关mRNA的内化显著增强。这一分析表明，细胞膜结合的初始步骤和随后的内化方式可能是最终蛋白表达的决定因素，而不是下游步骤。此外，我们比较了mRNA和pDNA在人脐静脉内皮细胞（HUVEC）中的转染效率，以证明mRNA传递的优势。
mRNA instead of DNA provides a new and attractive approach for gene therapy and genetic vaccination. Delivery of mRNA can bypass nuclear localization step enabling protein expression directly in cytoplasm through transcription. Current technologies for mRNA delivery are predominantly based on cationic liposomes with low activity for transfection. We, previously reported that applying inorganic nano-particles of carbonate apatite onto cationic liposome of DOTAP {N-[1-(2,3-dioleoloxy)propyl]-N,N,N-trimethyl ammonium chloride} resulted in high transfection potency for luciferase mRNA both in mitotic and non-mitotic cells. In this paper, we expanded the previous work and performed in detail study especially on two important parts, evaluating the image of the complex and analyzing the steps of gene delivery to detect the determinant factor for enhanced transfection potency. Transmission electron microscopic (TEM) observation clearly indicated the presence of inorganic carbonate apatite particles on mRNA–liposome complex and demonstrated the structure of the new hybrid carrier material. Due to apparently higher gravitational force of absorbed inorganic nano-particles, cellular contact and internalization of hybrid-particle-associated mRNA were significantly enhanced compared to DOTAP. This analysis indicates rather than downstream steps, initial steps of cell membrane binding and subsequent way of internalization could be the determinant factor for final protein expression. Moreover, we compared transfection efficiency of mRNA and pDNA in Human Umbilical Vein Endothelial cell (HUVEC) to demonstrate advantages of mRNA delivery.