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Development and in vitro and in vivo evaluation of gene delivery vectors based on dynein light chain Rp3 and synthetic peptides.

Grant number: 12/18850-8
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): March 01, 2013
Effective date (End): June 30, 2017
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Principal Investigator:Adriano Rodrigues Azzoni
Grantee:Marianna Teixeira de Pinho Favaro
Host Institution: Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Associated scholarship(s):15/20193-3 - Development of gene delivery vectors based on dynein light chain Rp3 and T22 peptide for CXCR4+-based cell targeting, BE.EP.DR


A major holdback to the wide application of gene therapy and DNA vaccination arises from the lack of an ideal vector, that is both safe and efficient. Although safer, nonviral vectors face a series of physical, enzymatic and diffusional barriers that limits the transgene's arrival to the nucleus of target cells. As a continuation of the work previously developed during the student's master's degree, the main objective of this project is to develop multifunctional nonviral vectors based on recombinant proteins, which can be evaluated in vitro as well as in vivo. We use the recombinant protein Rp3, a dynein light chain, aiming to explore the natural ability of molecular motors like dynein to carry cargoes from the periphery to inside the cell, through the microtubules network. Rp3 will be used fusioned to synthetic peptides: DNAbinding sequences, a cell-penetratin peptide (TAT) and a His Tag, and will be named T-Rp3. We expect to understand the mechanisms involved in the traffic of these vectors. By combining different techniques such as dynamic light scattering, zeta potential, interaction and transfection assays, as well as confocal microscopy, we will be able to correlate the physical-chemical properties of the pDNA:protein complex to the intracellular traffic and gene delivery efficiency. By transfecting mammalian cells we will be able to asses our vectors' efficiency to perform in vitro gene delivery, and finally, we will evaluate our vectors' potential through an in vivo model of cystic fibrosis (mice), performing immune histochemistry and quantification of repórter gene expression.

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Scientific publications
(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)
DE PINHO FAVARO, MARIANNA TEIXEIRA; UNZUETA, UGUTZ; DE CABO, MARTI; VILLAVERDE, ANTONIO; FERRER-MIRALLES, NEUS; AZZONI, ADRIANO RODRIGUES. Intracellular trafficking of a dynein-based nanoparticle designed for gene delivery. European Journal of Pharmaceutical Sciences, v. 112, p. 71-78, . (12/18850-8, 13/23780-1)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
FAVARO, Marianna Teixeira de Pinho. Evaluation of modular proteins for gene and drug delivery. 2017. Doctoral Thesis - Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia Campinas, SP.

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