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Cell-Penetrating Peptides for Transport of Plasmid DNA and microRNA: from Nanoscopic Structure to Gene Delivery.

Grant number: 16/24409-3
Support Opportunities: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
Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
Associated researchers:Cristiano Luis Pinto de Oliveira ; Ian William Hamley ; Sang Won Han ; Valeria Castelletto


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)

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Scientific publications (11)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MELLO, LUCAS R.; HAMLEY, IAN W.; MIRANDA, ANTONIO; ALVES, WENDEL A.; SILVA, EMERSON R.. beta-sheet assembly in amyloidogenic glutamic acid nanostructures: Insights from X-ray scattering and infrared nanospectroscopy. JOURNAL OF PEPTIDE SCIENCE, v. 25, n. 6, . (15/24018-1, 14/50867-3, 17/02317-2, 16/24409-3)
CASTELLETTO, VALERIA; EDWARDS-GAYLE, CHARLOTTE J. C.; HAMLEY, IAN W.; BARRETT, GLYN; SEITSONEN, JANI; RUOKOLAINEN, JANNE; DE MELLO, LUCAS RODRIGUES; DA SILVA, EMERSON RODRIGO. Model self-assembling arginine-based tripeptides show selective activity against Pseudomonas bacteria. CHEMICAL COMMUNICATIONS, v. 56, n. 4, p. 615-618, . (16/24409-3)
MELLO, LUCAS R.; AGUIAR, RODRIGO B.; YAMADA, RENATA Y.; MORAES, JANE Z.; HAMLEY, IAN W.; ALVES, WENDEL A.; REZA, MEHEDI; RUOKOLAINEN, JANNE; SILVA, EMERSON R.. Amphipathic design dictates self-assembly, cytotoxicity and cell uptake of arginine-rich surfactant-like peptides. JOURNAL OF MATERIALS CHEMISTRY B, v. 8, n. 12, p. 2495-2507, . (15/24018-1, 14/50867-3, 17/02317-2, 16/24409-3)
DE MELLO, LUCAS RODRIGUES; HAMLEY, IAN WILLIAM; CASTELLETTO, VALERIA; MORENO GARCI, BIANCA BONETTO; HAN, SANG WON; PINTO DE OLIVEIRA, CRISTIANO LUIS; DA SILV, EMERSON RODRIGO A. Nanoscopic Structure of Complexes Formed between DNA and the Cell-Penetrating Peptide Penetratin. Journal of Physical Chemistry B, v. 123, n. 42, p. 8861-8871, . (15/20206-8, 16/24409-3)
PELIN, JULIANE N. B. D.; GATTO, EMANUELA; VENANZI, MARIANO; CAVALIERI, FRANCESCA; OLIVEIRA, CRISTIANO L. P.; MARTINHO, HERCULANO; SILVA, EMERSON R.; AGUILAR, ANDREA M.; SOUZA, JULIANA S.; ALVES, WENDEL A.. Hybrid Conjugates Formed between Gold Nanoparticles and an Amyloidogenic Diphenylalanine-Cysteine Peptide. CHEMISTRYSELECT, v. 3, n. 24, p. 6756-6765, . (15/24018-1, 15/20446-9, 17/02317-2, 16/24409-3, 14/50867-3)
GERBELLI, BARBARA BIANCA; DA SILVA, EMERSON RODRIGO; SOARES, BRUNA MIRANDA; ALVES, WENDEL ANDRADE; DE OLIVEIRA, ELISABETH ANDREOLI. Multilamellar-to-Unilamellar Transition Induced by Diphenylalanine in Lipid Vesicles. Langmuir, v. 34, n. 5, p. 2171-2179, . (15/24018-1, 16/24409-3)
SOUZA, MARCIA I.; PRIETO, TATIANA; RODRIGUES, TIAGO; FERREIRA, FABIO F.; NASCIMENTO, FRANCISCO B.; RIBEIRO, ANDERSON O.; SILVA, EMERSON R.; GIUNTINI, FRANCESCA; ALVES, WENDEL A.. Conjugation with L, L-diphenylalanine Self-Assemblies Enhances In Vitro Antitumor Activity of Phthalocyanine Photosensitizer. SCIENTIFIC REPORTS, v. 7, . (15/24018-1, 14/50867-3, 14/50972-1, 16/24409-3)
PAZINATO, JULIA C. O.; VILLETTI, MARCOS A.; MERTINS, OMAR; SILVA, EMERSON R.; GARCIA, IRENE T. S.. Insights on Structuration of Peroxotungstic Acid in Aqueous Media. Journal of the Brazilian Chemical Society, v. 30, n. 4, p. 752-763, . (15/23948-5, 16/24409-3)
MELLO, LUCAS R.; HAMLEY, IAN W.; CASTELLETTO, VALERIA; GARCIA, BIANCA B. M.; LOURENCO, THIAGO C.; VASSILIADES, SANDRA V.; ALVES, WENDEL A.; HAN, SANG W.; SILVA, EMERSON R.. Self-assembly and intracellular delivery of DNA by a truncated fragment derived from the Trojan peptide Penetratin. SOFT MATTER, v. 16, n. 20, p. 4746-4755, . (14/50867-3, 17/02317-2, 16/24409-3, 15/24018-1)
FERNANDES PATTA, ANA C. M.; MATHEWS, PATRICK D.; MADRID, RAFAEL R. M.; RIGONI, VERA L. S.; SILVA, EMERSON R.; MERTINS, OMAR. Polyionic complexes of chitosan-N-arginine with alginate as pH responsive and mucoadhesive particles for oral drug delivery applications. International Journal of Biological Macromolecules, v. 148, p. 550-564, . (16/13368-4, 17/16722-6, 18/20482-3, 15/23948-5, 16/24409-3)
SILVA, EMERSON R.; LISTIK, EDUARDO; HAN, SANG W.; ALVES, WENDEL A.; SOARES, BRUNA M.; REZA, MEHEDI; RUOKOLAINEN, JANNE; HAMLEY, IAN W.. Sequence length dependence in arginine/phenylalanine oligopeptides: Implications for self-assembly and cytotoxicity. Biophysical Chemistry, v. 233, p. 1-12, . (16/24409-3, 15/24018-1)

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