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Nanoparticles formed by modular recombinant proteins: new platforms against tumor cells

Grant number: 20/00616-5
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): October 01, 2020
Effective date (End): July 31, 2022
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Adriano Rodrigues Azzoni
Grantee:Fernanda Santos de Oliveira
Host Institution: Escola Politécnica (EP). Universidade de São Paulo (USP). São Paulo , SP, Brazil


The low efficiency during the delivery of nucleic acids to tumor cells is a recurring problem in gene therapy studies for the treatment of cancer. This problem is mainly caused by the difficulty in directing and transporting the therapeutic molecules to the target cells due to the presence of numerous physical, enzymatic and diffusional barriers. Over the past few years, our research group has developed multifunctional recombinant proteins specially designed for gene delivery, seeking to mimic the ability of viruses to infect cells exploiting signaling, extra- and intra-cellular responses. The main objective of this project is the development and characterization of new multifunctional recombinant proteins capable of self-organizing into nanoparticles and efficiently transporting genetic material (small interfering RNA) into tumor cells. To accomplish this, the GFP reporter protein will be fused to a Desintegrin protein (Echistatin) that will serve as a targeting domain. The modular protein will also feature an arginine-based domain to nucleic acid interaction. The nanoparticles will be formed by combining siRNA and the modular proteins aiming to facilitate targeting, cell uptake and intracellular traffic of nucleic acids. We hope, therefore, to obtain RNAi-Protein nanoparticles capable of efficiently protect, and facilitate targeting, cell entry and intracellular trafficking of transgenes. The kinetics of nanoparticle formation, stability and physical-chemical parameters such as hydrodynamic diameter and zeta potential will then be evaluated and correlated with the efficiency of gene delivery for different types of tumor cells. For this, molecules of a model siRNA, capable of silencing the reporter gene Luciferase (Firefly Luciferase, Luc), will be used in the studies of nanoparticles formation and transfection. In this way, we hope to develop new vectors capable of efficiently transporting and directing transgenes to different tumor cell lines, as well as to obtain important information about the mechanisms involved in complexation, targeting, internalization and intracellular trafficking. (AU)

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