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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Intracellular trafficking of a dynein-based nanoparticle designed for gene delivery

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Autor(es):
de Pinho Favaro, Marianna Teixeira [1, 2] ; Unzueta, Ugutz [3, 4] ; de Cabo, Marti [5] ; Villaverde, Antonio [2, 3, 6] ; Ferrer-Miralles, Neus [2, 3, 6] ; Azzoni, Adriano Rodrigues [7]
Número total de Autores: 6
Afiliação do(s) autor(es):
[1] Univ Estadual Campinas, Ctr Biol Mol & Engn Genet, Av Candido Rondon 400, BR-13083875 Campinas, SP - Brazil
[2] Univ Autonoma Barcelona, Inst Biotecnol & Biomed, E-08193 Barcelona - Spain
[3] CIBER Bioingn Biomat & Nanomed CIBER BBN, Barcelona 08193 - Spain
[4] Hosp Santa Creu & Sant Pau, Inst Invest Biomed St Pau, Barcelona 08025 - Spain
[5] Univ Autonoma Barcelona, Serv Microscopia, E-08193 Barcelona - Spain
[6] Univ Autonoma Barcelona, Dept Genet & Microbiol, E-08193 Barcelona - Spain
[7] Univ Sao Paulo, Escola Politecn, Dept Engn Quim, Av Prof Luciano Gualberto, Trav 3, 380, BR-05508900 Sao Paulo, SP - Brazil
Número total de Afiliações: 7
Tipo de documento: Artigo Científico
Fonte: European Journal of Pharmaceutical Sciences; v. 112, p. 71-78, JAN 15 2018.
Citações Web of Science: 1
Resumo

The success of viruses in the delivery of the viral genome to target cells relies on the evolutionary selection of protein-based domains able to hijack the intermolecular interactions through which cells respond to intra-and extracellular stimuli. In an effort to mimic viral infection capabilities during non-viral gene delivery, a modular recombinant protein named T-Rp3 was recently developed, containing a DNA binding domain, a dynein molecular motor interacting domain, and a TAT-derived transduction domain. Here, we analyzed at the microscopic level the mechanisms behind the cell internalization and intracellular trafficking of this highly efficient modular protein vector. We found that the protein has the ability to self-assemble in discrete protein nanoparticles resembling viral capsids, to bind and condense plasmid DNA (pDNA), and to interact with eukaryotic cell membranes. Confocal and single particle tracking assays performed on living HeLa cells revealed that the T-Rp3 nanoparticles promoted an impressive speed of cellular uptake and perinuclear accumulation. Finally, the protein demonstrated to be a versatile vector, delivering siRNA at efficiencies comparable to Lipofectamine T. These results demonstrate the high potential of recombinant modular proteins with merging biological functions to fulfill several requirements needed to obtain cost-effective non-viral vectors for gene-based therapies. (AU)

Processo FAPESP: 13/23780-1 - Desenvolvimento de vetores não-virais de entrega gênica baseados na cadeia leve de Dineína Rp3
Beneficiário:Adriano Rodrigues Azzoni
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 12/18850-8 - Desenvolvimento e avaliação in vitro e in vivo de vetores de entrega gênica baseados na cadeia leve de Dineína Rp3 e peptídeos sintéticos
Beneficiário:Marianna Teixeira de Pinho Favaro
Linha de fomento: Bolsas no Brasil - Doutorado