Cobalt(II)-polybenzimidazole complexes as a nonviral gene carrier: effects of charges and benzimidazolyl groups.
The successful delivery of nucleic acids into cells and their availability at the intracellular sites of action remain a major challenge in gene therapy, because cells are resistant to the uptake of genetic materials and contain a variety of mechanisms that degrade nucleic acids. To develop a nonviral carrier, we have recently designed a series of metal complexes with polybenzimidazole (pbzim) that serve as DNA-condensing agents. In this study, we prepared a group of Co(II)-pbzim complexes that bear variable numbers of both positive charges and benzimidazole (bzim) groups. First, the formation of condensates at varied molar ratios of complex to DNA was examined by multiple biophysical measurements at pH 7.4. The results showed that the size, morphology, and surface charge of the condensates formed via a Co(II) complex dose-dependent process can be controlled by utilization of different complexes. Then, the imaging examination performed with both confocal and fluorescence microscope indicated that the DNA condensates can be internalized into cells. Luciferase activity assay showed that the DNA condensates formed at 1:1 of molar ratio are in favor of cell transfection because of their appropriate size, profile, and positive surface charges. The addition of the helper lipid DOPE can enhance both cellular uptake of the condensates and expression of the transferred genes. The MTT data indicated that cytotoxicity of both Co(II) complexes and condensates is enhanced with increasing positive charges and bzim groups. All the results revealed that the physical properties, cell transfection, and cytotoxicity of the condensates are regulated by the positive charges and bzim groups on the metal complexes.
Key Laboratory of Pesticide and Chemical Biology, Ministry of Education, and School of Chemistry, Central China Normal University, Wuhan 430079, China.
Rouwei
R
Jiang
Jun
J
Yin
Si
S
Hu
Xianggao
X
Meng
Changlin
C
Liu
D010582:Bacteriophage lambda / Q000235:genetics; D001562:Benzimidazoles / Q000737:chemistry* / Q000633:toxicity; D003035:Cobalt / Q000737:chemistry* / Q000633:toxicity; D056831:Coordination Complexes / Q000737:chemistry* / Q000633:toxicity; D004247:DNA / Q000737:chemistry* / Q000633:toxicity; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011108:Polymers / Q000737:chemistry* / Q000633:toxicity; D014162:Transfection / Q000379:methods*
D001562:Benzimidazoles; D056831:Coordination Complexes; D011108:Polymers; C549461:poly(benzimidazole); D003035:Cobalt; D004247:DNA