The effect of nocodazole on the transfection efficiency of lipid-bilayer coated gold nanoparticles
Nonviral vectors are safer than viral systems for gene therapy applications. However, the limited efficacy always prevents their being widely used in clinical practice. Aside from searching new gene nonviral vectors, many researchers focus on finding out new substances to improve the transfection efficiency of existent vectors. In this work, we found a transfection enhancer, nocodazole (NCZ), for dimethyldioctadecylammonium (DODAB, a cationic lipid) bilayer coated gold nanoparticles (AuNPs) mediated gene delivery. It was found that NCZ produces 3-fold transfection enhancement to HEK 293T cells assessed by flow cytometry (FCM). The result was further confirmed by luciferase assay, in which NCZ induced more than 5 times improvement in transfection efficiency after 48 h of transfection. The results from the inductively coupled plasma mass spectrometry (ICP-MS) and FCM showed that NCZ did not affect the internalization of DODAB–AuNPs/DNA complexes. The trafficking of the complexes by transmission electron microscopy (TEM) indicated that the interrupted transportation of the complexes to the lysosomes contributed greatly to the transfection enhancement. Therefore, NCZ can be used as a transfection enhancer in DODAB–AuNPs mediated transfection system. This work also gave an insight to improving the efficiency of lipid-mediated transfection: modifying lipid on gold nanoparticles and pre-treating cells by NCZ before the transfection.
Journal: Biomaterials - Volume 30, Issue 7, March 2009, Pages 1382–1388