Studying a possible functional circuitry involving microRNAs, MITF, MYO5A/DLC2, RA...
| Grant number: | 16/24409-3 |
| Support type: | Regular Research Grants |
| Duration: | April 01, 2017 - March 31, 2019 |
| Field of knowledge: | Biological Sciences - Biophysics - Molecular Biophysics |
| Principal Investigator: | Emerson Rodrigo da Silva |
| Grantee: | Emerson Rodrigo da Silva |
| Home Institution: | Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil |
| Assoc. researchers: | Cristiano Luis Pinto de Oliveira ; Ian William Hamley ; Sang Won Han ; Valeria Castelletto |
Abstract
In the late 1980's, researches related to HIV conducted to the surprising discovery that certain protein domains are able to cross eukaryotic membranes. These short sequences, usually containing between 5 and 40 amino acids, were quickly called cell-penetrating peptides (CPPs) and their potential for designing synthetic vectors suitable for gene therapy was immediately recognized. Currently, scientific literature reports on more than 150 CPPs which are efficient to transport bioactive compounds; however, detailed data on their nanoscopic organization are still very scarce. This scenario leads to a lack of fundamental information for establishing structure/transfection correlations, making difficulty optimizing these transporters. In this Project, we will contribute to fill this gap through systematic investigations on the nanoscopic ordering of conjugates formulated between CPPs and nucleic acids. We will study complexes involving either long circular plasmid DNA (4.7 kb) DNA or small RNA (20-22 nt). The nanoscopic organization will be elucidated through a sophisticated combination of biophysical techniques including X-ray and neutron small-angle scattering, fiber X-ray diffraction, electron and atomic force microscopy and spectroscopic approaches. Concomitantly, transfection assays will be carried out to investigate either expression or suppression of green fluorescent protein into mammal cells in order to unveil the role of structure on efficiency of these vectors. As a final goal, we aim to identify structural patterns common to different classes of CPPs and, from these universal features, design new structurally-optimized CPPs for transfection. (AU)