RESUMEN
Gene therapy has arisen as a pioneering technique to treat diseases by direct employment of nucleic acids as medicine. The major historical problem is to develop efficient and safe systems for the delivery of therapeutic genes into the target cells. Carbon nanotubes (CNTs) have demonstrated considerable promise as delivery vectors due to their (i) high aspect ratio and (ii) capacity to translocate through plasma membranes, known as the nanoneedle effect. To leverage these advantages, close attention needs to be paid to the physicochemical characteristics of the CNTs used. CNTs with different diameters (thinner and thicker) were treated by chemical oxidation to produce shorter fragments. Rigid (thick) and flexible (thin) CNTs, and their shortened versions, were coated with polyallylamine (ppAA) by plasma-enhanced chemical vapor deposition. The ppAA coating leads to a positively charged CNT surface that is able to electrostatically bind the green fluorescent protein plasmid reporter. This study shows how rigidity and length can affect their (i) behavior in biological media, (ii) ability to transfect in vitro, and (iii) biodistribution in vivo. This study also generates a set of basic design rules for the development of more efficient CNT-based gene-delivery vectors.
Asunto(s)
Nanotubos de Carbono , Técnicas de Transferencia de Gen , Terapia Genética , Plásmidos , Distribución TisularRESUMEN
The surface modification of carbon nanotubes (CNTs) can be tailored to allow the formation of a complex between these potential carriers with DNA. In this chapter, protocols developed in our lab to prepare transfection vectors through the modification of MWCNTs are described. The protocol includes sections focused on the reduction of CNTs length, protocols to increase the dispersability of CNTs and finally, protocols for surface modification to attach through electrostatic interactions DNA to the CNTs.