Nanotechnology is currently used in the development of new strategies for the release of genes and therapies against cancer. One of the aims of gene therapy is to introduce a copy of a given gene into the defective cells of a patient to restore an impaired biological function. The principle of gene transfer is based on the use of DNA complexing agents known as vectors, natural or synthetic, capable of protecting the genetic material from degradation in biological fluids and able to transport it through cell membranes for its integration into the transcription mechanism of a target cell. Cationic polymers that interact electrostatically with negatively charged DNA allow the formation of positive polyplexes capable of transporting and releasing DNA into cells. Currently, there are different physicochemical and biophysical techniques that allow characterizing the nanoparticles resulting from the compaction of DNA with the different available polycations. Dynamic light scattering (DLS) allows accessing to the particle size and the polydispersity index; conductivity measurements allow obtaining information about the stoichiometry of the nanoparticle formation; electrophoretic mobility measurements give information about the net charge of the particles; TEM and AFM microscopy show the morphology and dimensions; agarose gels provide information on DNA/polycation interactions, degradation by DNases, serum; among other techniques (1). The selection of the vector to be used to transfer the DNA, together with an adequate formulation, are key parameters that allow an optimal transfection efficiency to be obtained.
Topics of the short course:
DAY 1: Introduction to natural polymers and biopolymers, main characteristics of biopolymers, water-soluble polyelectrolyte properties, main characteristics of non-viral vectors for gene therapy.
DAY 2: Polyplexes from electrostatic interactions, biopolymers/DNA nanoparticles formulation, complexation mechanism (stoichiometry, net charge evolution), main characteristics of the nanoparticles.
References:
[1] L. M. Bravo-Anaya, J.F. A. Soltero, M. Rinaudo, International Journal of Biological Macromolecules 88 (2016) 345–353.
