Sabouri-Rad Sarvenaz, Oskuee Reza Kazemi, Mahmoodi Asma, Gholami Leila, Malaekeh-Nikouei Bizhan
School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Targeted Drug Delivery Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Bioimpacts. 2017;7(3):139-145. doi: 10.15171/bi.2017.17. Epub 2017 Aug 16.
Cationic polymers have the potential to be modified to achieve an ideal gene vector lacking viral vector defects. The aim of the present study was to improve polyallylamine (PAA) transfection efficiency and to reduce cytotoxicity by incorporating of cell-penetrating peptides (CPPs). To prepare the peptide-based polyplexes, PAA (15 kDa) was modified with 2 peptides (TAT and CyLoP-1) by covering the 0.5% and 1% of amines. Buffer capacity and DNA condensation ability of modified polymer, particle size and zeta potential of nanoparticles, cell viability, and transfection activity of vectors were evaluated. In low carrier to plasmid (C/P) weight ratios such as 0.5 and 1, the unmodified polymer was more capable to condense the DNA compared to the synthesized vectors. In C/P ratio of 2, the plasmid was fully condensed in all vectors. The size of polyplexes ranged from 195 to 240 nm. The zeta potential was almost as the same as PAA and varied from 25 to 27 mV. All polyplexes increased the buffer capacity compared to PAA. The transfection efficiency was improved compared to unmodified polymer especially in the vectors modified with 1% of TAT or CyLoP-1 peptides in C/P ratio of 2. The cytotoxicity of prepared vectors was less than PAA. In most ratios, the cytotoxicity of the CyLoP-1 modified samples was less than the TAT modified ones. Modification of PAA with CPPs improved the transfection activity of vector.
阳离子聚合物有被修饰以获得一种缺乏病毒载体缺陷的理想基因载体的潜力。本研究的目的是通过掺入细胞穿透肽（CPPs）来提高聚烯丙胺（PAA）的转染效率并降低细胞毒性。为了制备基于肽的多聚体，用两种肽（TAT和CyLoP-1）修饰15 kDa的PAA，覆盖0.5%和1%的胺基。评估了修饰聚合物的缓冲能力和DNA凝聚能力、纳米颗粒的粒径和zeta电位、细胞活力以及载体的转染活性。在低载体与质粒（C/P）重量比（如0.5和1）下，与合成载体相比，未修饰的聚合物更能凝聚DNA。在C/P比为2时，质粒在所有载体中都完全凝聚。多聚体的大小在195至240 nm之间。zeta电位几乎与PAA相同，在25至27 mV之间变化。与PAA相比，所有多聚体都提高了缓冲能力。与未修饰的聚合物相比，转染效率得到了提高，特别是在C/P比为2时用1%的TAT或CyLoP-1肽修饰的载体中。制备的载体的细胞毒性小于PAA。在大多数比例下，CyLoP-1修饰样品的细胞毒性小于TAT修饰的样品。用CPPs修饰PAA提高了载体的转染活性。
