Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Int J Pharm. 2012 May 10;427(2):393-9. doi: 10.1016/j.ijpharm.2012.01.062. Epub 2012 Feb 8.
Peptide based gene carriers are among the most promising non-viral vectors for gene delivery to eukaryotic cells. We have engineered a new fusion peptide using recombinant technology with the purpose of overcoming the cell barriers to gene delivery. A His- tagged multi-domain peptide was expressed in Escherichia coli BL21 (DE3) pLysS and purified using Ni-NTA resin. The fusion peptide is composed of two repeats of truncated histone H1 peptide to condense pDNA, a fusogenic peptide to disrupt endosome membranes and a nuclear localization signal to enhance translocation of pDNA towards nucleus. The results demonstrated that the vector can effectively condense plasmid DNA into nanoparticles with average sizes of 200 nm. The fusogenic peptide in the vector structure also showed membrane disruptive effect in the endosomal pH. Overall, the transfection efficiency of the vector demonstrated that it holds great promise as a nontoxic and effective non-viral gene carrier.
基于肽的基因载体是最有前途的非病毒载体之一，可将基因递送至真核细胞。我们使用重组技术设计了一种新的融合肽，旨在克服基因传递的细胞障碍。带有 His 标签的多功能肽在大肠杆菌 BL21（DE3）pLysS 中表达，并使用 Ni-NTA 树脂进行纯化。融合肽由两个截断的组蛋白 H1 肽重复序列组成，用于浓缩 pDNA，一个融合肽用于破坏内涵体膜，一个核定位信号用于增强 pDNA 向核内的转移。结果表明，该载体可以有效地将质粒 DNA 浓缩成平均大小为 200nm 的纳米颗粒。载体结构中的融合肽在内涵体 pH 值下也表现出膜破坏效应。总的来说，该载体的转染效率表明，它作为一种无毒有效的非病毒基因载体具有很大的应用潜力。
