Majidi Asia, Nikkhah Maryam, Sadeghian Faranak, Hosseinkhani Saman
Department of Nanobiotechnology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran.
Eur J Pharm Biopharm. 2016 Oct;107:191-204. doi: 10.1016/j.ejpb.2016.06.017. Epub 2016 Jun 28.
In last decades great efforts have been devoted to the study of development of recombinant peptide based vectors that consist of biological motifs with potential applications in gene therapy. Recombinant Biomimetic Chimeric Vectors (rBCVs) are biopolymeric nanocarriers that are designed to mimic viral features to overcome the cellular obstacles in gene transferring pathway into cell nucleus. In this research, we designed and genetically engineered three novel rBCVs with similar sequences that differed in motifs arrangement and motif abundance: MPG-2H1, 2TMPG-2H1 and 2RMPG-2H1. The MPG as a famous amphipathic cell penetrating peptide is the main segment of these constructs which was studied for the first time in association with truncated histone H1 DNA condensing motif. Through the performance of several physicochemical and biological assays, the rBCVs were remarkably examined regarding transfection efficiency. The main objective of this study is focused on the importance of motif design in transfection efficiency of rBCVs on one hand, and the assessment of correlation between structural features and functionality of motifs on the other hand. The results revealed that all three kinds of rBCVs/pDNA nanoparticles with average sizes of 200nm could overwhelm the cellular obstacles associated with gene transfer, and lead to efficient gene delivery. Furthermore, no significant toxicity was perceived and efficient endosome disruptive activity was obtained. It is noteworthy to say among three mentioned constructs 2RMPG-2H1 showed the highest transfection efficiency. Overall the peptide based vectors hold great promise as a nontoxic and effective gene carrier in vitro and in vivo, besides the rational design possibility as the most vital advantages over the other non-viral gene delivery vectors.
在过去几十年中，人们致力于研究基于重组肽的载体的开发，这些载体由具有基因治疗潜在应用的生物基序组成。重组仿生嵌合载体（rBCV）是生物聚合物纳米载体，旨在模仿病毒特征，以克服基因转移至细胞核过程中的细胞障碍。在本研究中，我们设计并基因工程改造了三种具有相似序列但基序排列和基序丰度不同的新型rBCV：MPG-2H1、2TMPG-2H1和2RMPG-2H1。MPG作为一种著名的两亲性细胞穿透肽，是这些构建体的主要部分，首次与截短的组蛋白H1 DNA凝聚基序联合进行研究。通过进行多种物理化学和生物学检测，对rBCV的转染效率进行了显著检测。本研究的主要目标一方面集中在基序设计对rBCV转染效率的重要性，另一方面集中在基序的结构特征与功能之间相关性的评估。结果表明，所有三种平均粒径为200nm的rBCV/pDNA纳米颗粒都能克服与基因转移相关的细胞障碍，并实现高效的基因递送。此外，未观察到明显的毒性，且获得了有效的内体破坏活性。值得注意的是，在上述三种构建体中，2RMPG-2H1表现出最高的转染效率。总体而言，基于肽的载体作为一种无毒且有效的基因载体在体外和体内具有巨大潜力，此外，与其他非病毒基因递送载体相比，其合理设计的可能性是最重要的优势。
