Zinc(?)-polybenzimidazole Complexes As Nonviral Gene Carriers

Gene therapy is to put human normal genes or therapeutic genes into human target cells through a certain way to correct genetic defects or to play a role in treatment,so as to achieve the purpose of the treatment of diseases of the new technology.With the development of medicine,gene therapy as a new means of treating disease,has been paid much attention of scientific workers.In the course of the study of gene therapy,how to carry gene into the target cells for expression is one of the major problems in urgent need of scientific research to solve,which requires safe and efficient gene targeting vector.A series of multi benzimidazole metal complexes of our research group has the property of molecular aggregation induced by DNA,and through the experiment confirmed that the complexes induced DNA molecules aggregate to membrane into the cell,and found that the complexes as non viral gene vectors with the transfection efficiency.In this work,we synthesized ten kinds of multi benzimidazole zinc metal complexes,and systematic study of the properties of these ten complexes as gene carrier,and by the addition of cell penetrating peptides and polyethylene glycol to form multiple different complexes to improve the transfection efficiency of the complexes.We use multi benzimidazole ligands with different?IDB,NTB,EDTB,CTB?and three kinds of zinc metal salts?ZnCl₂,Zn?NO₃?₂·6H₂O,Zn?ClO₄?₂·6H₂O?ten with different numbers of benzimidazole synthesis(2,3,4,and He Zheng charge?+1?+2?,the benzene and imidazole zinc complexes.Firstly,through a combination of complexes with DNA UV absorption spectra,found that the complexes can bind to DNA,and its binding ability increased with the number of benzimidazole and positive charge number.Using agarose gel electrophoresis retardation test?EMSA?on the properties of complexes to DNA condensation,and further through the TEM?TEM?investigated concentration and structure on DNA condensation,found that different reaction conditions can affect the aggregate particle size and morphology,and the structures of the complexes also have a great influence on the properties of condensed matter.Therefore,by choosing the appropriate DNA complexes induced aggregation,can be compact and ordered nanometer spherical agglomerates.The experiment confirmed by luciferase and green fluorescent protein expression,complexes/DNA condensate can be released into the cell expression plasmid.By MTT method to detect the coagulation of cell toxicity,the study found that such complexes with DNA to form agglomerate body with low cell toxicity.In a word,can play as non viral gene vectors gene vector function of these metal complexes.Additionally,We study the effect of cell penetrating peptide?CPP?and polyethylene glycol?PEG?for the cell transfection efficiency,as well as the research of transmembrane pathway.Through the experiment has proved that the cell penetrating peptide and polyethylene glycol can enhance significantly transfection efficiency.The transmembrane pathway was confirmed by the experimental results of coagulation membrane endocytic pathway by clathrin mediated.